U.S. patent number 11,028,089 [Application Number 16/343,102] was granted by the patent office on 2021-06-08 for pyrimido-diazepinone kinase scaffold compounds and methods of treating dclk1/2-mediated disorders.
This patent grant is currently assigned to DANA-FARBER CANCER INSTITUTE, INC.. The grantee listed for this patent is DANA-FARBER CANCER INSTITUTE, INC.. Invention is credited to Fleur M. Ferguson, Nathanael S. Gray.
United States Patent |
11,028,089 |
Ferguson , et al. |
June 8, 2021 |
Pyrimido-diazepinone kinase scaffold compounds and methods of
treating DCLK1/2-mediated disorders
Abstract
The present invention relates to use of pyrimido-diazepinone
compounds that are able to modulate protein kinases such as
doublecortin-like kinase (DCLK1) and doublecortin-like kinase 2
(DCLK2), which are members of serine/threonine-protein kinase
family and Ca.sup.2+/calmodulin-dependent protein kinase class of
enzymes, and the use of such compounds in the treatment of various
diseases, disorders or conditions.
Inventors: |
Ferguson; Fleur M. (Cambridge,
MA), Gray; Nathanael S. (Boston, MA) |
Applicant: |
Name |
City |
State |
Country |
Type |
DANA-FARBER CANCER INSTITUTE, INC. |
Boston |
MA |
US |
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Assignee: |
DANA-FARBER CANCER INSTITUTE,
INC. (Boston, MA)
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Family
ID: |
1000005602712 |
Appl.
No.: |
16/343,102 |
Filed: |
October 18, 2017 |
PCT
Filed: |
October 18, 2017 |
PCT No.: |
PCT/US2017/057126 |
371(c)(1),(2),(4) Date: |
April 18, 2019 |
PCT
Pub. No.: |
WO2018/075608 |
PCT
Pub. Date: |
April 26, 2018 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20190315753 A1 |
Oct 17, 2019 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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62409457 |
Oct 18, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P
35/00 (20180101); C07D 487/04 (20130101) |
Current International
Class: |
A61K
31/551 (20060101); C07D 487/04 (20060101); A61P
35/00 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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3424929 |
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Sep 2019 |
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EP |
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WO 2014/160430 |
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Oct 2014 |
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WO |
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2015/117083 |
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Aug 2015 |
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WO |
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2015/117087 |
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Aug 2015 |
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WO |
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Other References
Patel et al., Biochemical and Structural Insights Into
Doublecortin-like Kinase Domain 1, Structure, vol. 24, No. 9, pp.
1550-1561. (Year: 2016). cited by examiner .
Ferguson et al., Discovery of a Series of
5,11-Dihydro-6H-Benzo[e]]pyrimido[5,4-b][1,4]diazepin-6-ones as
Selective PI3K-delta/gamma Inhibitiors, ACS Medicinal Chemistry
Letters, vol. 7, No. 10, pp. 908-912. (Year: 2016). cited by
examiner .
Lin et al., ERK5 Kinase Activity is Not Required for Cellular
Immune Response, bioRxiv, Immunology, p. 38513/1-38513/47. (Year:
2016). cited by examiner .
Kanhed et al., Benzo[e]pyrimido[5,4-b][1,4]diazepin-6(11H)-one
Derivatives as Aurora A Kinase Inhibitors: LQTA-QSAR Analysis and
Detailed Systematic Validation of the Developed Model, Molecular
Diversity, vol. 19, No. 4, pp. 965-974. (Year: 2015). cited by
examiner .
Munoz et al., Optimisation of LRRK2 Inhibitors and Assessment of
Functional Efficacy in Cell-Based Models of Neuroinflammation,
European Journal of Medicinal Chemistry, vol. 95, pp. 29-34. (Year:
2015). cited by examiner .
Weygant et al., Small Molecule Kinase Inhibitor LRRK2-IN-1
Demonstrates Potent Activity Against Colorectal and Pancreatic
Cancer Through Inhibition of Doublecortin-Like Kinase 1, Molecular
Cancer, vol. 13, p. 103/1-103/14. (Year: 2014). cited by examiner
.
Sureban et al., XMD8-92 Inhibits Pancreatic Tumor Xenograft Growth
via a DCLK1-Dependent Mechanism, Cancer Letters, vol. 351, No. 1,
pp. 151-161 (Year: 2014). cited by examiner .
Leurman et al., Phosphoproteomic Evaluation of Pharmacological
Inhibition of Leucine-Rich Repeat Kinase 2 Reveals Significient
Off-Target Effects of LRRK-2-IN-1, Journal of Neurochemistry, vol.
128, No. 4, pp. 561-576. (Year: 2014). cited by examiner .
Deng et al., Structural Determinants For ERK5 (MAPK7) and Leucine
Rich Repeat Kinase 2 Activities of Benzo[e]pyrimido-[5,4-b]
diazepine-6(11H)-ones, European Journal of Medicinal Chemistry,
vol. 70, pp. 758-767. (Year: 2013). cited by examiner .
Elkins et al., X-Ray Crystal Structure of ERK5 (MAPK7) in Complex
with a Specific Inhibitor, Journal of Medicinal Chemistry, vol. 56
, No. 11, pp. 4413-4421. (Year: 2013). cited by examiner .
Kwiatkowski et al., Selective Aurora Kinase Inhibitors Identified
Using a Taxol-lnduced Checkpoint Sensitivity Screen, ACS Chemical
Biology, vol. 7, No. 1, pp. 185-196. (Year: 2012). cited by
examiner .
Miduturu et al., High-Throughput Kinase Profiling: A More Efficient
Approach Toward the Discovery of New Kinase Inhibitors, Chemistry
& Biology, vol. 18, No. 7, pp. 868-879. (Year: 2011). cited by
examiner .
Deng et al., Discovery of a
Benzo[e]pyrimido-[5,4-b][1,4]diazepin-6(11H)-one as a Potent and
Selective Inhibitor of Big MAP Kinase 1, ACS Medicinal Chemistry
Letters, vol. 2, No. 3, pp. 195-200. (Year: 2011). cited by
examiner .
Ferguson, Fleur M., et al., "Discovery of a Selective Inhibitor of
Doublecortin Like Kinase 1", Nat. Chem. Biol. Apr. 6, 2020, doi:
10.1038/s41589-020-0506-0. cited by applicant .
Ito, Hiromitsu, et al., "Dominant Expression of DCLK1 in Human
Pancreatic Cancer Stem Cells Accelerates Tumor Invasion and
Metastasis", PLOS One, 2016, vol. 11, No. 1, e0146564. cited by
applicant .
Westphalen, C. Benedikt, et al., "Long-Lived Intestinal Tuft Cells
Serve as Colon Cancer-Initiating Cells", J. Clin. Invest., 2014,
vol. 124, No. 3, pp. 1283-1295. cited by applicant .
Weygant, Nathaniel, et al., "Small Molecule Kinase Inhibitor
LRRK2-IN-1 Demonstrates Potent Activity Against Colorectal and
Pancreatic Cancer Through Inhibition of Doublecortin-like Kinase
1", Mol. Cancer, 2014, vol. 13, No. 103. cited by
applicant.
|
Primary Examiner: Coleman; Brenda L
Attorney, Agent or Firm: Burns & Levinson, LLP Clarke;
Daniel W.
Parent Case Text
RELATED APPLICATIONS
This application is a national stage application, filed under 35
U.S.C. .sctn. 371, of International Application No.
PCT/US2017/057126, filed Oct. 18, 2017, which claims the benefit of
priority under 35 U.S.C. .sctn. 119(e) to U.S. Provisional
Application No. 62/409,457, filed Oct. 18, 2016, each of which is
incorporated herein by reference in its entirety.
Claims
What is claimed:
1. A method of treating pancreatic cancer in a subject, the method
comprising administering to the subject a compound of formula F-1:
##STR00097## or a pharmaceutically acceptable salt thereof,
wherein, R.sub.1 is ##STR00098## R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3; R.sub.5 is hydrogen
or methyl; and R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and p is 0-4.
2. The method of claim 1, wherein the compound is of formula F-1-a:
##STR00099## or a pharmaceutically acceptable salt thereof.
3. The method of claim 1, wherein the subject is administered
simultaneously or sequentially an additional therapeutic agent,
wherein the additional therapeutic agent comprises an
anti-inflammatory agent, which is azathioprine, cyclophosphamide,
or sulfasalazine, or a chemotherapeutic agent, which is imatinib
mesylate, adriamycin, dexamethasone, vincristine, cyclophosphamide,
fluorouracil, topotecan, taxol, an interferon, or a platinum
derivative.
4. The method of claim 1, wherein the subject is a human.
5. A compound of formula F-1: ##STR00100## or a pharmaceutically
acceptable salt thereof, wherein, R.sub.1 is ##STR00101## R.sub.2
is --CH.sub.2--CH.sub.2F, --CH.sub.2--CHF.sub.2, or
--CH.sub.2--CF.sub.3; R.sub.5 is hydrogen or methyl; and R.sub.6 is
hydrogen or optionally substituted alkyl; each R.sub.7 is
independently alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and p is 0-4.
6. The compound of claim 5, wherein the compound is of formula
F-1-a: ##STR00102## or a pharmaceutically acceptable salt
thereof.
7. A pharmaceutical composition comprising the compound of claim 5,
or a pharmaceutically acceptable salt thereof.
8. The method of claim 1, wherein the compound is ##STR00103## or a
pharmaceutically acceptable salt thereof.
9. The compound of claim 5, wherein the compound is ##STR00104## or
a pharmaceutically acceptable salt thereof.
10. A compound which is: ##STR00105## ##STR00106## ##STR00107## or
a pharmaceutically acceptable salt thereof.
11. A method of treating pancreatic cancer in a subject, the method
comprising administering to the subject a compound of claim 10, or
a pharmaceutically acceptable salt thereof.
Description
TECHNICAL FIELD
The present invention relates to pyrimido-diazepinone compounds
which are able to modulate protein kinases such as
doublecortin-like kinase 1 (DCLK1) and doublecortin-like kinase 2
(DCLK2), which are members of serine/threonine-protein kinase
family and Ca.sup.2+/calmodulin-dependent protein kinase class of
enzymes, and the use of such compounds in the treatment of various
diseases, disorders or conditions.
BACKGROUND OF THE INVENTION
Protein kinases represent a large family of proteins, which play a
central role in the regulation of a wide variety of cellular
processes and maintaining control over cellular function. Protein
kinases constitute a large family of structurally related enzymes
that are responsible for the control of a variety of signal
transduction processes within the cell (see Hardie, G and Hanks, S.
The Protein Kinase Facts Book, I and II, Academic Press, San Diego,
Calif.: 1995). Protein kinases are thought to have evolved from a
common ancestral gene due to the conservation of their structure
and catalytic function. Almost all kinases contain a similar
250-300 amino acid catalytic domain. The kinases may be categorized
into families by the substrates they phosphorylate (e.g.,
protein-tyrosine, protein-serine/threonine, lipids etc).
In general, protein kinases mediate intracellular signaling by
catalyzing a phosphoryl transfer from a nucleoside triphosphate to
a protein acceptor that is involved in a signaling pathway. These
phosphorylation events act as molecular on/off switches that can
modulate or regulate the target protein biological function. These
phosphorylation events are ultimately triggered in response to a
variety of extracellular and other stimuli. Examples of such
stimuli include environmental and chemical stress signals (e.g.,
shock, heat shock, ultraviolet radiation, bacterial endotoxin, and
H2O2), cytokines (e.g., interleukin-1 (IL-I) and tumor necrosis
factor alpha (TNF-a), and growth factors (e.g., granulocyte
macrophage-colony stimulating factor (GM-CSF), and fibroblast
growth factor (FGF)). An extracellular stimulus may affect one or
more cellular responses related to cell growth, migration,
differentiation, secretion of hormones, activation of transcription
factors, muscle contraction, glucose metabolism, control of protein
synthesis, survival and regulation of the cell cycle.
Described herein are compounds that inhibit the activity of one or
more isoforms of the protein kinase DCLK1/2 and are, therefore,
expected to be useful in the treatment of kinase-associated
diseases.
SUMMARY OF THE INVENTION
In one aspect, the invention provides a method of treating a
disease mediated by a kinase such as doublecortin-like kinase 1
(DCLK1) and/or doublecortin-like kinase 2 (DCLK2). The method
comprises administering of a kinase inhibitor compound, e.g., a
compound of formula A:
##STR00001##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
X is CHR.sub.4, CR.sub.4, NH, NR.sub.4 or N;
Y is NR.sub.5, N, S, SO, SO.sub.2, O, CHR.sub.5, or CR.sub.5;
wherein at least one of X and Y is NH, NR.sub.4, NR.sub.5, N, S,
SO, SO.sub.2, or O;
A is a single bond or double bond;
B is a single bond or double bond, wherein both A and B are not
double bonds;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.4 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.5 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
or R.sub.3 and X, together with the atoms to which they are
attached, form a 3-8 membered carbocyclic, aryl, heterocyclic, or
heteroaryl; each of which is optionally substituted;
or X and Y, together with the atoms to which they are attached,
form a 3-8 membered carbocyclic, aryl, heterocyclic, or heteroaryl;
each of which is optionally substituted; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In one aspect, the method of treating a disease in a subject
mediated by doublecortin-like kinase (DCLK1) and/or DCLK2 comprises
administering to the subject a compound of formula F-1:
##STR00002##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.5 is hydrogen, optionally substituted alkyl, optionally
substituted aralkyl, or optionally substituted carbocyclic; and
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In some embodiments, R.sub.5 is methyl.
In some embodiments, R.sub.2 is unsubstituted alkyl.
In some embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen.
In some embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In some embodiments, the compound is of formula F-1-a:
##STR00003##
or a pharmaceutically acceptable salt, ester or prodrug
thereof.
In embodiments, R.sub.1 is methyl, ethyl, propyl, iso-propyl,
butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl, piperidinyl,
pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridizinyl, quinolinyl, thienyl, thiazolyl, oxazolyl,
isoxazolyl, pyrrolyl, furanyl, isoquinolinyl, imiazolyl, or
triazolyl, each of which may be optionally substituted.
In embodiments, R.sub.1 is phenyl or pyridyl, each of which may be
optionally substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, and carbocyclic,
each of which may be further substituted; wherein each R.sub.A is
independently selected from alkyl, alkenyl, carbocyclic, aryl,
heteroaryl, and heterocyclic, or two R.sub.A on the same atom
combine to form a heterocyclic, each of which may be further
substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from alkoxy, CO.sub.2Me,
##STR00004##
In embodiments, R.sub.1 is phenyl, pyridyl, pyrimidinyl, furyl,
pyrrolyl, pyrazolyl, imidazolyl, thienyl, or
bicyclo[1.1.1]pent-1-yl, each of which may be optionally
substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, carbocyclic,
SO.sub.2(R.sub.A), SO.sub.3 (R.sub.A), SO.sub.2N(R.sub.A)
(R.sub.A), SO.sub.2NH(R.sub.A), SO.sub.2NH.sub.2,
PO(OR.sub.A)(OR.sub.A), or PO(OR.sub.A)(R.sub.A), each of which may
be further substituted; and each R.sub.A is independently selected
from alkyl, alkenyl, carbocyclic, aryl, heteroaryl, and
heterocyclic, or two R.sub.A on the same atom combine to form a
heterocyclic, each of which may be further substituted.
In some embodiments, R.sub.1 is selected from the group consisting
of:
##STR00005##
In embodiments, the compound inhibits DCLK1 and/or DCLK2.
In embodiments, the disease is cancer or a proliferation
disease.
In some embodiments, the disease is lung, colon, breast, prostate,
liver, pancreas, brain, kidney, ovaries, stomach, skin, and bone
cancers, gastric, breast, pancreatic cancer, glioma, and
hepatocellular carcinoma, papillary renal carcinoma, head and neck
squamous cell carcinoma, leukemias, lymphomas, myelomas, solid
tumors, or blood-borne cancers (e.g., chronic lymphocytic leukemia
(CLL), follicular lymphoma (FL), or indolent non-Hodgkin's lymphoma
(iNHL).
In some embodiments, the disease is Barretts' esophagus, esophageal
cancer, salivary gland malignancies, colon and colorectal cancer,
intestinal cancer, gastric cancer, pancreatic cancer, skin cancer
or neuroblastoma.
In embodiments, the disease is a liver disease.
In some embodiments, the disease is a fatty liver disease,
non-alcoholic fatty liver disease (NAFLD), non-alcoholic
steatohepatitis (NASH), cirrhosis, fatty liver disease resulting
from hepatitis, fatty liver disease resulting from obesity, fatty
liver disease resulting from diabetes, fatty liver disease
resulting from insulin resistance, fatty liver disease resulting
from hypertriglyceridemia, Abetalipoproteinemia, glycogen storage
diseases, Wolmans disease, or acute fatty liver of pregnancy.
In embodiments, the disease is a neurodegenerative disease.
In some embodiments, the disease is Alzheimer's disease (AD),
Parkinson's disease (PD), Huntington's (HD) diseases, amyotrophic
lateral sclerosis (ALS), spinal muscular atrophy (SMA),
schizophrenia, attention-deficit/hyperactivity disorder (ADHD),
fetal alcohol syndrome and diabetic encephalopathy.
In embodiments, the subject is administered an additional
therapeutic agent.
In embodiments, said additional therapeutic agent are administered
simultaneously or sequentially.
In embodiments, said additional therapeutic agent is a
chemotherapeutic agent.
In one aspect, a method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprises contacting a cell with a
compound of formula F-1:
##STR00006##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.5 is hydrogen, optionally substituted alkyl, optionally
substituted aralkyl, or optionally substituted carbocyclic; and
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment comprises administering a compound of formula F-1:
##STR00007##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.5 is hydrogen, optionally substituted alkyl, optionally
substituted aralkyl, or optionally substituted carbocyclic; and
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In embodiments, the subject is a human.
In embodiments, the compound has a Ki for inhibiting the
doublecortin-like kinase (DCLK1/2) less than about 1 .mu.M, less
than about 500 nM, less than about 100 nM, less than about 50 nM,
less than about 40 nM, less than about 30 nM, less than about 20
nM, or less than about 15 nM.
In some embodiments, R.sub.5 is methyl.
In some embodiments, R.sub.2 is unsubstituted alkyl.
In some embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen.
In some embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In embodiments, the compound is of formula F-1-a:
##STR00008##
or a pharmaceutically acceptable salt, ester or prodrug
thereof.
In embodiments, R.sub.1 is methyl, ethyl, propyl, iso-propyl,
butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl, piperidinyl,
pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridizinyl, quinolinyl, thienyl, thiazolyl, oxazolyl,
isoxazolyl, pyrrolyl, furanyl, isoquinolinyl, imiazolyl, or
triazolyl, each of which may be optionally substituted.
In embodiments, R.sub.1 is phenyl or pyridyl, each of which may be
optionally substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, and carbocyclic,
each of which may be further substituted;
wherein each R.sub.A is independently selected from alkyl, alkenyl,
carbocyclic, aryl, heteroaryl, and heterocyclic, or two R.sub.A on
the same atom combine to form a heterocyclic, each of which may be
further substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from alkoxy, CO.sub.2Me,
##STR00009##
In embodiments, R.sub.1 is phenyl, pyridyl, pyrimidinyl, furyl,
pyrrolyl, pyrazolyl, imidazolyl, thienyl, or
bicyclo[1.1.1]pent-1-yl, each of which may be optionally
substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, carbocyclic,
SO.sub.2(R.sub.A), SO.sub.3(R.sub.A), S.sub.2N(R.sub.A) (R.sub.A),
SO.sub.2NH(R.sub.A), SO.sub.2NH.sub.2, PO(OR.sub.A)(OR.sub.A), or
PO(OR.sub.A)(R.sub.A), each of which may be further substituted;
and
wherein each R.sub.A is independently selected from alkyl, alkenyl,
carbocyclic, aryl, heteroaryl, and heterocyclic, or two R.sub.A on
the same atom combine to form a heterocyclic, each of which may be
further substituted.
In some embodiments, R.sub.1 is selected from the group consisting
of:
##STR00010##
In one aspect, the method of treating a disease in a subject
mediated by a kinase such as doublecortin-like kinase (DCLK1/2)
comprises administering to the subject a compound of formula
A-1:
##STR00011##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
X is CHR.sub.4, CR.sub.4, NH, NR.sub.4 or N;
Y is NR.sub.5, N, S, SO, SO.sub.2, O, CHR.sub.5, or CR.sub.5;
wherein at least one of X and Y is NH, NR.sub.4, NR.sub.5, N, S,
SO, SO.sub.2, or O;
A is a single bond or double bond;
B is a single bond or double bond, wherein both A and B are not
double bonds;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.4 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.5 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
or R.sub.3 and X, together with the atoms to which they are
attached, form a 3-8 membered carbocyclic, aryl, heterocyclic, or
heteroaryl; each of which is optionally substituted;
or X and Y, together with the atoms to which they are attached,
form a 3-8 membered carbocyclic, aryl, heterocyclic, or heteroaryl;
each of which is optionally substituted; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula A-1, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula A-1, or a
pharmaceutically acceptable salt, ester or prodrug thereof.
In embodiments, the compound has a structure according to formula
B-1:
##STR00012##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is aryl, or heteroaryl, wherein R.sub.1 may be optionally
substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen or methyl;
R.sub.4 is hydrogen or methyl; and
R.sub.6 is hydrogen.
In embodiments, the compound has a structure according to formula
C-1:
##STR00013##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is aryl, heteroaryl, which may be optionally
substituted;
R.sub.2 is hydrogen or methyl;
R.sub.3 is hydrogen;
R.sub.4 is hydrogen; and
R.sub.6 is hydrogen.
In embodiments, the compound has a structure according to formula
D-1:
##STR00014##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-6.
In embodiments, the compound has a structure according to formula
E-1:
##STR00015##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen or optionally substituted alkyl;
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-6.
In embodiments, the compound has a structure according to formula
F-I:
##STR00016##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
Y is S, SO, SO.sub.2, N, or O;
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In embodiments, the compound has a structure according to formula
G-1:
##STR00017##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or 3
heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic, wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.5 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In one aspect, the method of treating a disease in a subject
mediated by doublecortin-like kinase (DCLK1) and/or
doublecortin-like kinase 2 (DCLK2) comprises administering to the
subject a compound of formula I-2:
##STR00018##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
A is a single bond or double bond;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen, optionally substituted alkyl, optionally
substituted cycloalkyl, and optionally substituted heterocyclyl;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula I-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula I-2, or a
pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the method of treating a disease in a subject
mediated by doublecortin-like kinase (DCLK1) and/or
doublecortin-like kinase 2 (DCLK2) comprises comprising
administering to the subject a compound of formula II-2:
##STR00019##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
E is NR.sub.2 or CHR.sub.2;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula II-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula II-2, or
a pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the method of treating a disease in a subject
wherein the disease is mediated by doublecortin-like kinase (DCLK1)
and/or doublecortin-like kinase 2 (DCLK2), the method comprising
administering to the subject a compound of formula III-2:
##STR00020##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula III-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula III-2, or
a pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the method of treating a disease in a subject
wherein the disease is mediated by doublecortin-like kinase (DCLK1)
and/or doublecortin-like kinase 2 (DCLK2), the method comprising
administering to the subject a compound of formula IV-2:
##STR00021##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula IV-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula IV-2, or
a pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, method of treating a disease in a subject mediated
by doublecortin-like kinase (DCLK1) and/or doublecortin-like kinase
2 (DCLK2), the method comprising administering to the subject a
compound of formula V-2:
##STR00022##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
X is an optional substituent as defined for formula I;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is --OH or --O-(optionally substituted alkyl);
R.sub.4 is hydrogen or optionally substituted alkyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula V-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula V-2, or a
pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the method of treating a disease in a subject
mediated by doublecortin-like kinase (DCLK1) and/or
doublecortin-like kinase 2 (DCLK2), the method comprising
administering to the subject a compound of formula VI-2:
##STR00023## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl; or
two X moieties on adjacent atoms of the thiophene ring can form,
together with the atoms to which they are attached, a phenyl ring;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula VI-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula VI-2, or
a pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the method of treating a disease in a subject
wherein the disease is mediated by doublecortin-like kinase (DCLK1)
and/or doublecortin-like kinase 2 (DCLK2), the method comprising
administering to the subject a compound of formula VII-2:
##STR00024## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen, optionally substituted alkyl, optionally
substituted cycloalkyl, and optionally substituted heterocyclyl;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula VII-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula VII-2, or
a pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the method of treating a disease in a subject
mediated by doublecortin-like kinase (DCLK1) and/or
doublecortin-like kinase 2 (DCLK2), the method comprising
administering to the subject a compound of formula VIII-2:
##STR00025## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
Z is O or S;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen, optionally substituted alkyl, optionally
substituted cycloalkyl, and optionally substituted heterocyclyl;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula VIII-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula VIII-2,
or a pharmaceutically acceptable salt, ester or prodrug
thereof.
In one aspect, the method of treating a disease in a subject
mediated by doublecortin-like kinase (DCLK1) and/or
doublecortin-like kinase 2 (DCLK2), the method comprising
administering to the subject a compound of formula IX-2:
##STR00026## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
A is a single bond or double bond;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
Y is hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, and optionally substituted heterocyclyl;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 and R.sub.2' are each independently hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl;
or Y and R.sub.2' can form, together with the atoms to which they
are attached, a five-membered ring; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In another aspect, the method for reducing doublecortin-like kinase
(DCLK1/2)-dependent cell growth comprising contacting a cell with a
compound of formula IX-2, or a pharmaceutically acceptable salt,
ester or prodrug thereof.
In another aspect, the method of inhibiting doublecortin-like
kinase (DCLK1/2) in a subject identified as in need of such
treatment, comprising administering a compound of formula IX-2, or
a pharmaceutically acceptable salt, ester or prodrug thereof.
In one aspect, the present invention provides a compound of formula
F-1:
##STR00027##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.5 is hydrogen, optionally substituted alkyl, optionally
substituted aralkyl, or optionally substituted carbocyclic; and
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In embodiments, R.sub.5 is methyl.
In embodiments, R.sub.2 is unsubstituted alkyl.
In some embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen.
In some embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In embodiments, the compound is of formula F-1-a:
##STR00028##
or a pharmaceutically acceptable salt, ester or prodrug
thereof.
In embodiments, R.sub.2 is unsubstituted alkyl.
In some embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen.
In some embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In embodiments, R.sub.1 is methyl, ethyl, propyl, iso-propyl,
butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl, piperidinyl,
pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridizinyl, quinolinyl, thienyl, thiazolyl, oxazolyl,
isoxazolyl, pyrrolyl, furanyl, isoquinolinyl, imiazolyl, or
triazolyl, each of which may be optionally substituted.
In some embodiments, R.sub.1 is phenyl or pyridyl, each of which
may be optionally substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, and carbocyclic,
each of which may be further substituted;
wherein each R.sub.A is independently selected from alkyl, alkenyl,
carbocyclic, aryl, heteroaryl, and heterocyclic, or two R.sub.A on
the same atom combine to form a heterocyclic, each of which may be
further substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from alkoxy, CO.sub.2Me,
##STR00029##
In embodiments, R.sub.1 is phenyl, pyridyl, pyrimidinyl, furyl,
pyrrolyl, pyrazolyl, imidazolyl, thienyl, or
bicyclo[1.1.1]pent-1-yl, each of which may be optionally
substituted.
In some embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, carbocyclic,
SO.sub.2(R.sub.A), SO.sub.3 (R.sub.A), SO.sub.2N(R.sub.A)
(R.sub.A), SO.sub.2NH(R.sub.A), SO.sub.2NH.sub.2,
PO(OR.sub.A)(OR.sub.A), or PO(OR.sub.A)(R.sub.A), each of which may
be further substituted; and
wherein each R.sub.A is independently selected from alkyl, alkenyl,
carbocyclic, aryl, heteroaryl, and heterocyclic, or two R.sub.A on
the same atom combine to form a heterocyclic, each of which may be
further substituted.
In some embodiments, R.sub.1 is selected from the group consisting
of:
##STR00030##
In embodiments, the compound of formula F-1-a inhibits DCLK1 and/or
DCLK2.
Further provided in the present invention is a pharmaceutical
composition comprising the compound (e.g. formula F-1, formula
F-1-a, formula A-1 and etc.) as described herein, or a
pharmaceutically acceptable salt, ester or prodrug thereof.
Additionally, in one aspect, the intention provides methods of
treating a disease in a subject, wherein the disease is mediated by
doublecortin-like kinase (DCLK1) and/or doublecortin-like kinase 2
(DCLK2), the method comprising administering to the subject the
compound as described herein, or a pharmaceutically acceptable
salt, ester or prodrug thereof.
In another aspect, the invention provides methods for reducing
doublecortin-like kinase (DCLK1/2)-dependent cell growth comprising
contacting a cell with the compound as described herein, or a
pharmaceutically acceptable salt, ester or prodrug thereof.
In another aspect, the invention provides methods of inhibiting
doublecortin-like kinase (DCLK1/2) in a subject identified as in
need of such treatment, comprising administering the compound as
described herein, or a pharmaceutically acceptable salt, ester or
prodrug thereof.
In one aspect, the invention features a method of treating a
disease in a subject mediated by a kinase that is doublecortin-like
kinase (DCLK1/2) comprising administering to the subject a compound
as described herein (e.g., a compound of formula F-1, formula F-1-a
or formula A-1), or a pharmaceutically acceptable salt, ester or
prodrug thereof.
In another aspect, the invention features a method for reducing
doublecortin-like kinase (DCLK1/2)-dependent cell growth comprising
contacting a cell with a compound as described herein (e.g., a
compound of formula F-1, formula F-1-a or formula A-1) or a
pharmaceutically acceptable salt, ester or prodrug thereof.
In another aspect, the invention features a method of inhibiting
doublecortin-like kinase (DCLK1/2) in a subject identified as in
need of such treatment, comprising administering a compound as
described (e.g., a compound of formula F-1, formula F-1-a or
formula A-1), or a pharmaceutically acceptable salt, ester or
prodrug thereof.
In another aspect, the invention provides a method for reducing
kinase-dependent cell growth comprising contacting a cell with a
kinase inhibitor compound as described herein or a pharmaceutically
acceptable ester, salt, or prodrug thereof.
In other aspects, the invention provides a method of inhibiting
kinase in a subject identified as in need of such treatment,
comprising administering a kinase inhibitor compound as described
herein, or a pharmaceutically acceptable ester, salt, or prodrug
thereof.
In one aspect, the invention provides a kit comprising a compound
capable of inhibiting kinase activity selected from one or more
kinase inhibitor compounds described herein, or a pharmaceutically
acceptable ester, salt, or prodrug thereof, and instructions for
use in treating cancer.
In one aspect, the invention provides a pharmaceutical composition
comprising a kinase inhibitor compound as described herein, or a
pharmaceutically acceptable ester, salt, or prodrug thereof,
together with a pharmaceutically acceptable carrier.
In one aspect, the invention provides a method of synthesizing a
kinase inhibitor compound as described herein.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 illustrates selectivity data generated using KINOMEscan.RTM.
platform for Compound 2 at 1 uM concentration and this image was
generated using TREEspot.TM. Software Tool.
FIG. 2 shows inhibition profiles (IC.sub.50) of compounds of the
invention against DCLK1.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
Listed below are definitions of various terms used to describe this
invention. These definitions apply to the terms as they are used
throughout this specification and claims, unless otherwise limited
in specific instances, either individually or as part of a larger
group.
The term "alkyl," as used herein, refers to saturated, straight- or
branched-chain hydrocarbon radicals containing, in certain
embodiments, between one and six, or one and eight carbon atoms,
respectively. Examples of C.sub.1-C.sub.6 alkyl radicals include,
but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl,
tert-butyl, neopentyl, n-hexyl radicals; and examples of
C.sub.1-C.sub.8 alkyl radicals include, but are not limited to,
methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl,
n-hexyl, heptyl, octyl radicals.
The term "alkenyl," as used herein, denotes a monovalent group
derived from a hydrocarbon moiety containing, in certain
embodiments, from two to six, or two to eight carbon atoms having
at least one carbon-carbon double bond. The double bond may or may
not be the point of attachment to another group. Alkenyl groups
include, but are not limited to, for example, ethenyl, propenyl,
butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.
The term "alkynyl," as used herein, denotes a monovalent group
derived from a hydrocarbon moiety containing, in certain
embodiments, from two to six, or two to eight carbon atoms having
at least one carbon-carbon triple bond. The alkynyl group may or
may not be the point of attachment to another group. Representative
alkynyl groups include, but are not limited to, for example,
ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl and the like.
The term "alkoxy" refers to an --O-alkyl radical.
The term "aryl," as used herein, refers to a mono- or poly-cyclic
carbocyclic ring system having one or more aromatic rings, fused or
non-fused, including, but not limited to, phenyl, naphthyl,
tetrahydronaphthyl, indanyl, idenyl and the like.
The term "aralkyl," as used herein, refers to an alkyl residue
attached to an aryl ring. Examples include, but are not limited to,
benzyl, phenethyl and the like.
The term "cycloalkyl," as used herein, denotes a monovalent group
derived from a monocyclic or polycyclic saturated or partially
unsatured carbocyclic ring compound. Examples of
C.sub.3-C.sub.8-cycloalkyl include, but not limited to,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl and
cyclooctyl; and examples of C.sub.3-C.sub.12-cycloalkyl include,
but not limited to, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2] octyl. Also
contemplated are a monovalent group derived from a monocyclic or
polycyclic carbocyclic ring compound having at least one
carbon-carbon double bond by the removal of a single hydrogen atom.
Examples of such groups include, but are not limited to,
cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl,
cycloheptenyl, cyclooctenyl, and the like.
The term "heteroaryl," as used herein, refers to a mono- or
poly-cyclic (e.g., bi-, or tri-cyclic or more) fused or non-fused,
radical or ring system having at least one aromatic ring, having
from five to ten ring atoms of which one ring atoms is selected
from S, O and N; zero, one or two ring atoms are additional
heteroatoms independently selected from S, O and N; and the
remaining ring atoms are carbon. Heteroaryl includes, but is not
limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,
imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl,
oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl,
benzimidazolyl, benzooxazolyl, quinoxalinyl, and the like.
The term "heteroaralkyl," as used herein, refers to an alkyl
residue attached to a heteroaryl ring. Examples include, but are
not limited to, pyridinylmethyl, pyrimidinylethyl and the like.
The term "heterocycloalkyl," as used herein, refers to a
non-aromatic 3-, 4-, 5-, 6- or 7-membered ring or a bi- or
tri-cyclic group fused of non-fused system, where (i) each ring
contains between one and three heteroatoms independently selected
from oxygen, sulfur and nitrogen, (ii) each 5-membered ring has 0
to 1 double bonds and each 6-membered ring has 0 to 2 double bonds,
(iii) the nitrogen and sulfur heteroatoms may optionally be
oxidized, (iv) the nitrogen heteroatom may optionally be
quaternized, and (iv) any of the above rings may be fused to a
benzene ring. Representative heterocycloalkyl groups include, but
are not limited to, [1,3]dioxolane, pyrrolidinyl, pyrazolinyl,
pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,
piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl,
thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl.
The term "alkylamino" refers to a group having the structure
--NH(C.sub.1-C.sub.12 alkyl) where C.sub.1-C.sub.12 alkyl is as
previously defined.
The term "acyl" includes residues derived from acids, including but
not limited to carboxylic acids, carbamic acids, carbonic acids,
sulfonic acids, and phosphorous acids. Examples include aliphatic
carbonyls, aromatic carbonyls, aliphatic sulfonyls, aromatic
sulfinyls, aliphatic sulfinyls, aromatic phosphates and aliphatic
phosphates. Examples of aliphatic carbonyls include, but are not
limited to, acetyl, propionyl, 2-fluoroacetyl, butyryl, 2-hydroxy
acetyl, and the like. In accordance with the invention, any of the
aryls, substituted aryls, heteroaryls and substituted heteroaryls
described herein, can be any aromatic group. Aromatic groups can be
substituted or unsubstituted.
The terms "halo" and "halogen," as used herein, refer to an atom
selected from fluorine, chlorine, bromine and iodine.
As described herein, compounds of the invention may optionally be
substituted with one or more substituents, such as are illustrated
generally above, or as exemplified by particular classes,
subclasses, and species of the invention. It will be appreciated
that the phrase "optionally substituted" is used interchangeably
with the phrase "substituted or unsubstituted." In general, the
term "substituted", whether preceded by the term "optionally" or
not, refers to the replacement of hydrogen radicals in a given
structure with the radical of a specified substituent. Unless
otherwise indicated, an optionally substituted group may have a
substituent at each substitutable position of the group, and when
more than one position in any given structure may be substituted
with more than one substituent selected from a specified group, the
substituent may be either the same or different at every position.
The terms "optionally substituted", "optionally substituted alkyl,"
"optionally substituted "optionally substituted alkenyl,"
"optionally substituted alkynyl", "optionally substituted
cycloalkyl," "optionally substituted cycloalkenyl," "optionally
substituted aryl", "optionally substituted heteroaryl," "optionally
substituted aralkyl", "optionally substituted heteroaralkyl,"
"optionally substituted heterocycloalkyl," and any other optionally
substituted group as used herein, refer to groups that are
substituted or unsubstituted by independent replacement of one,
two, or three or more of the hydrogen atoms thereon with
substituents including, but not limited to:
--F, --Cl, --Br, --I,
--OH, protected hydroxy,
--NO.sub.2, --CN,
--NH.sub.2, protected amino, --NH--C.sub.1-C.sub.12-alkyl,
--NH--C.sub.2-C.sub.12-alkenyl, --NH--C.sub.2-C.sub.12-alkenyl,
--NH--C.sub.3-C.sub.12-cycloalkyl, --NH-aryl, --NH-heteroaryl,
--NH-heterocycloalkyl, -dialkylamino, -diarylamino,
-diheteroarylamino,
--O--C.sub.1-C.sub.12-alkyl, --O--C.sub.2-C.sub.12-alkenyl,
--O--C.sub.2-C.sub.12-alkenyl, --O--C.sub.3-C.sub.12-cycloalkyl,
--O-aryl, --O-heteroaryl, --O-heterocycloalkyl,
--C(O)-- C.sub.1-C.sub.12-alkyl, --C(O)-- C.sub.2-C.sub.12-alkenyl,
--C(O)-- C.sub.2-C.sub.12-alkenyl,
--C(O)--C.sub.3-C.sub.12-cycloalkyl, --C(O)-aryl,
--C(O)-heteroaryl, --C(O)-heterocycloalkyl,
--CONH.sub.2, --CONH--C.sub.1-C.sub.12-alkyl,
--CONH--C.sub.2-C.sub.12-alkenyl, --CONH--C.sub.2-C.sub.12-alkenyl,
--CONH--C.sub.3-C.sub.12-cycloalkyl, --CONH-aryl,
--CONH-heteroaryl, --CONH-- heterocycloalkyl,
--OCO.sub.2-- C.sub.1-C.sub.12-alkyl,
--OCO.sub.2--C.sub.2-C.sub.12-alkenyl, --OCO.sub.2--
C.sub.2-C.sub.12-alkenyl, --OCO.sub.2--C.sub.3-C.sub.12-cycloalkyl,
--OCO.sub.2-aryl, --OCO2-heteroaryl, --OCO.sub.2-heterocycloalkyl,
--OCONH.sub.2, --OCONH--C.sub.1-C.sub.12-alkyl,
--OCONH--C.sub.2-C.sub.12-alkenyl,
--OCONH--C.sub.2-C.sub.12-alkenyl,
--OCONH--C.sub.3-C.sub.12-cycloalkyl, --OCONH-- aryl,
--OCONH-heteroaryl, --OCONH-heterocycloalkyl,
--NHC(O)-- C.sub.1-C.sub.12-alkyl,
--NHC(O)--C.sub.2-C.sub.12-alkenyl,
--NHC(O)--C.sub.2-C.sub.12-alkenyl,
--NHC(O)--C.sub.3-C.sub.12-cycloalkyl, --NHC(O)-aryl,
--NHC(O)-heteroaryl, --NHC(O)-- heterocycloalkyl, --NHCO.sub.2--
C.sub.1-C.sub.12-alkyl, --NHCO.sub.2-- C.sub.2-C.sub.12-alkenyl,
--NHCO.sub.2-- C.sub.2-C.sub.12-alkenyl, --NHCO.sub.2--
C.sub.3-C.sub.12-cycloalkyl, --NHCO.sub.2-- aryl,
--NHCO.sub.2-heteroaryl, --NHCO.sub.2-heterocycloalkyl,
--NHC(O)NH.sub.2, --NHC(O)NH--C.sub.1-C.sub.12-alkyl,
--NHC(O)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(O)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(O)NH--C.sub.3-C.sub.12-cycloalkyl, --NHC(O)NH-aryl,
--NHC(O)NH-heteroaryl, --NHC(O)NH-heterocycloalkyl, NHC(S)NH.sub.2,
--NHC(S)NH--C.sub.1-C.sub.12-alkyl,
--NHC(S)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(S)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(S)NH--C.sub.3-C.sub.12-cycloalkyl, --NHC(S)NH-aryl,
--NHC(S)NH-heteroaryl, --NHC(S)NH-heterocycloalkyl,
--NHC(NH)NH.sub.2, --NHC(NH)NH--C.sub.1-C.sub.12-alkyl,
--NHC(NH)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)NH--C.sub.3-C.sub.12-cycloalkyl, --NHC(NH)NH-aryl,
--NHC(NH)NH-heteroaryl, --NHC(NH)NH-- heterocycloalkyl,
--NHC(NH)--C.sub.1-C.sub.12-alkyl,
--NHC(NH)--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)--C.sub.3-C.sub.12-cycloalkyl, --NHC(NH)-aryl, --NHC(NH)--
heteroaryl, --NHC(NH)-heterocycloalkyl,
--C(NH)NH--C.sub.1-C.sub.12-alkyl,
--C(NH)NH--C.sub.2-C.sub.12-alkenyl,
--C(NH)NH--C.sub.2-C.sub.12-alkenyl,
--C(NH)NH--C.sub.3-C.sub.12-cycloalkyl, --C(NH)NH-aryl,
--C(NH)NH-heteroaryl, --C(NH)NH-heterocycloalkyl,
--S(O)--C.sub.1-C.sub.12-alkyl, --S(O)--C.sub.2-C.sub.12-alkenyl,
--S(O)--C.sub.2-C.sub.12-alkenyl,
--S(O)--C.sub.3-C.sub.12-cycloalkyl, --S(O)-aryl,
--S(O)-heteroaryl, --S(O)-heterocycloalkyl --SO.sub.2NH.sub.2,
--SO.sub.2NH--C.sub.1-C.sub.12-alkyl,
--SO.sub.2NH--C.sub.2-C.sub.12-alkenyl,
--SO.sub.2NH--C.sub.2-C.sub.12-alkenyl,
--SO.sub.2NH--C.sub.3-C.sub.12-cycloalkyl, --SO.sub.2NH-aryl,
--SO.sub.2NH-heteroaryl, --SO.sub.2NH-heterocycloalkyl,
--NHSO.sub.2--C.sub.1-C.sub.12-alkyl,
--NHSO.sub.2--C.sub.2-C.sub.12-alkenyl,
--NHSO.sub.2--C.sub.2-C.sub.12-alkenyl,
--NHSO.sub.2--C.sub.3-C.sub.12-cycloalkyl, --NHSO.sub.2-aryl,
--NHSO.sub.2-heteroaryl, --NHSO.sub.2-heterocycloalkyl,
--CH.sub.2NH.sub.2, --CH.sub.2SO.sub.2CH.sub.3, -aryl, -arylalkyl,
-heteroaryl, -heteroarylalkyl, -heterocycloalkyl,
--C.sub.3-C.sub.12-cycloalkyl, polyalkoxyalkyl, polyalkoxy,
-methoxymethoxy, -methoxyethoxy, --SH, --S--C.sub.1-C.sub.12-alkyl,
--S--C.sub.2-C.sub.12-alkenyl, --S--C.sub.2-C.sub.12-alkenyl,
--S--C.sub.3-C.sub.12-cycloalkyl, --S-aryl, --S-heteroaryl,
--S-heterocycloalkyl, or methylthiomethyl.
It is understood that the aryls, heteroaryls, alkyls, and the like
can be further substituted.
The term "cancer" includes, but is not limited to, the following
cancers: epidermoid Oral: buccal cavity, lip, tongue, mouth,
pharynx; Cardiac: sarcoma (angiosarcoma, fibrosarcoma,
rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma,
lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell or
epidermoid, undifferentiated small cell, undifferentiated large
cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial
adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;
Gastrointestinal: esophagus (squamous cell carcinoma, larynx,
adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma,
lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma,
insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma),
small bowel or small intestines (adenocarcinoma, lymphoma,
carcinoid tumors, Karposi's sarcoma, leiomyoma, hemangioma, lipoma,
neurofibroma, fibroma), large bowel or large intestines
(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,
leiomyoma), colon, colon-rectum, colorectal; rectum, Genitourinary
tract: kidney (adenocarcinoma, WiIm's tumor [nephroblastoma],
lymphoma, leukemia), bladder and urethra (squamous cell carcinoma,
transitional cell carcinoma, adenocarcinoma), prostate
(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal
carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial
cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma);
Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma,
hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma,
biliary passages; Bone: osteogenic sarcoma (osteosarcoma),
fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma,
Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma),
multiple myeloma, malignant giant cell tumor chordoma,
osteochronfroma (osteocartilaginous exostoses), benign chondroma,
chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell
tumors; Nervous system: skull (osteoma, hemangioma, granuloma,
xanthoma, osteitis deformans), meninges (meningioma,
meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,
glioma, ependymoma, germinoma [pinealoma], glioblastoma multiform,
oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),
spinal cord neurofibroma, meningioma, glioma, sarcoma);
Gynecological: uterus (endometrial carcinoma), cervix (cervical
carcinoma, pre-tumor cervical dysplasia), ovaries (ovarian
carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma,
unclassified carcinoma], granulosa-thecal cell tumors,
Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma),
vulva (squamous cell carcinoma, intraepithelial carcinoma,
adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell
carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal
rhabdomyosarcoma), fallopian tubes (carcinoma), breast;
Hematologic: blood (myeloid leukemia [acute and chronic], acute
lymphoblastic leukemia, chronic lymphocytic leukemia,
myeloproliferative diseases, multiple myeloma, myelodysplastic
syndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignant
lymphoma] hairy cell; lymphoid disorders; Skin: malignant melanoma,
basal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma,
keratoacanthoma, moles dysplastic nevi, lipoma, angioma,
dermatofibroma, keloids, psoriasis, Thyroid gland: papillary
thyroid carcinoma, follicular thyroid carcinoma; medullary thyroid
carcinoma, undifferentiated thyroid cancer, multiple endocrine
neoplasia type 2A, multiple endocrine neoplasia type 2B, familial
medullary thyroid cancer, pheochromocytoma, paraganglioma; Adrenal
glands: neuroblastoma, and blood-borne cancers such as chronic
lymphocytic leukemia (CLL), follicular lymphoma (FL) and indolent
non-Hodgkin's lymphoma (iNHL).
Thus, the term "cancerous cell" as provided herein, includes a cell
afflicted by any one of the above-identified conditions.
The term "liver disease" is damage to or disease of the liver that
can lead to failure of liver functions. The term "liver disease"
includes, but is not limited to, the following diseases: hepatitis,
alcoholic liver disease, hereditary diseases such as
hemochromatosis and Wilson's disease, a fatty liver disease,
non-alcoholic fatty liver disease (NAFLD), non-alcoholic
steatohepatitis (NASH), cirrhosis, primary biliary cirrhosis,
primary sclerosing cholangitis, Budd-Chiari syndrome, fatty liver
disease resulting from hepatitis, fatty liver disease resulting
from obesity, fatty liver disease resulting from diabetes, fatty
liver disease resulting from insulin resistance, fatty liver
disease resulting from hypertriglyceridemia, Abetalipoproteinemia,
glycogen storage diseases, Wolmans disease, acute fatty liver of
pregnancy, Weber-Christian disease, Gilber's syndrome, Wolmans
disease, acute fatty liver of pregnancy, or lipodystrophy.
In embodiments, the disease is a neurodegenerative disease.
In some embodiments, the disease is Alzheimer's disease (AD),
Parkinson's disease (PD), Huntington's (HD) diseases, amyotrophic
lateral sclerosis (ALS), spinal muscular atrophy (SMA),
schizophrenia, attention-deficit/hyperactivity disorder (ADHD),
fetal alcohol syndrome and diabetic encephalopathy.
The term "Kinase Panel" is a list of kinases including, but not
limited to, MPS1 (TTK), ERK5 (BMK1, MAPK7), polo kinase 1,2,3, or
4, Ack1, Ack2, Abl, DCAMKL1, ABL1, Abl mutants, DCAMKL2, ARK5, BRK,
MKNK2, FGFR4, TNK1, PLK1, ULK2, PLK4, PRKD1, PRKD2, PRKD3, ROS1,
RPS6KA6, TAOK1, TAOK3, TNK2, Bcr-Abl, GAK, cSrc, TPR-Met, Tie2,
MET, FGFR3, Aurora, Axl, Bmx, BTK, c-kit, CHK2, Flt3, MST2, p70S6K,
PDGFR, PKB, PKC, Raf, ROCK-H, Rsk1, SGK, TrkA, TrkB, TrkC, AAK1,
ABL1, ABL1(E255K), ABL1(F317I), ABL1(F317L), ABL1(H396P),
ABL1(M351T), ABL1(Q252H), ABL1(T315I), ABL1(Y253F), ABL2, ACVR1,
ACVR1B, ACVR2A, ACVR2B, ACVRL1, ADCK3, ADCK4, AKT1, AKT2, AKT3,
ALK, AMPK-alpha1, AMPK-alpha2, ANKK1, ARK5, ASK1, ASK2, AURKA,
AURKB, AURKC, AXL, BIKE, BLK, BMPR1A, BMPR1B, BMPR2, BMX, BRAF,
BRAF(V600E), BRK, BRSK1, BRSK2, BTK, CAMK1, CAMK1D, CAMK1G, CAMK2A,
CAMK2D, CAMK2G, CAMK4, CAMKK1, CAMKK2, CDC.sub.2L1, CDC.sub.2L2,
CDK11, CDK2, CDK3, CDK5, CDK7, CDK8, CDK9, CDKL2, CDKL3, CDKL5,
CHECK1, CHEK2, CIT, CLK1, CLK2, CLK3, CLK4, CSF1R, CSK, CSNK1A1L,
CSNK1D, CSNK1E, CSNK1G1, CSNK1G3, CSNK2A1, CSNK2A2, CTK, DAPK1,
DAPK2, DAPK3, DCAMKL1 (DLCK1), DCAMKL2 (DCLK2), DCAMKL3, DDR1,
DDR2, DLK, DMPK, DMPK2, DRAK1, DRAK2, DYRK1A, DYRK1B, DYRK2, EGFR,
EGFR (E746-A750DEL), EGFR (G719C), EGFR (G719S), EGFR(L747-E749del,
A750P), EGFR(L747-S752del, P753S), EGFR(L747-T751del,Sins),
EGFR(L858R), EGFR(L858R,T790M), EGFR(L861Q), EGFR(S752-1759del),
EPHAL EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHB1,
EPHB2, EPHB3, EPHB4, EPHB6, ERBB2, ERBB3, ERBB4, ERK1, ERK2, ERK3,
ERK4, ERK5, ERK8, ERN1, FAK, FER, FES, FGFR1, FGFR2, FGFR3,
FGFR3(G697C), FGFR4, FGR, FLT1, FLT3, FLT3(D835H), FLT3(D835Y),
FLT3(ITD), FLT3(K663Q), FLT3(N841I), FLT4, FRK, FYN, GAK,
GCN2(Kin.Dom.2,S808G), GRK1, GRK4, GRK7, GSK3A, GSK3B, HCK, HIPK1,
HIPK2, HIPK3, HIPK4, HPK1, HUNK, ICK, IGF1R, IKK-ALPHA, IKK-BETA,
IKK-EPSILON, INSR, INSRR, IRAK1, IRAK3, ITK,
JAK1(JH1domain-catalytic), JAK1(JH2domain-pseudokinase),
JAK2(JH1domain-catalytic), JAK3(JH1domain-catalytic), JNK1, JNK2,
JNK3, KIT, KIT(D816V), KIT(L576P), KIT(V559D), KIT(V559D,T670I),
KIT(V559D,V654A), LATS1, LATS2, LCK, LIMK1, LIMK2, LKB1, LOK, LTK,
LYN, LZK, MAK, MAP3K1, MAP2K15, MAP3K2, MAP3K3, MAP3K4, MAP4K2,
MAP4K3, MAP4K5, MAPKAPK2, MAPKAPK5, MARK1, MARK2, MARK3, MARK4,
MAST1, MEK1, MEK2, MEK3, MEK4, MEK6, MELK, MERTK, MET, MET(M1250T),
MET(Y1235D), MINK, MKNK1, MKNK2, MLCK, MLK1, MLK2, MLK3, MRCKA,
MRCKB, MST1, MST1R, MST2, MST3, MST4, MUSK, MYLK, MYLK2, MYO3A,
MYO3B, NDR1, NDR2, NEK1, NEK2, NEK5, NEK6, NEK7, NEK9, NIM1, NLK,
OSR1, p38-alpha, p38-beta, p38-delta, p38-gamma, PAK1, PAK2, PAK3,
PAK4, PAK6, PAK7, PCTK1, PCTK2, PCTK3, PDGFRA, PDGFRB, PDPK1,
PFTAIRE2, PFTK1, PHKG1, PHKG2, PIK3C2B, PIK3C2G, PIK3CA,
PIK3CA(C420R), PIK3CA(E542K), PIK3CA(E545A), PIK3CA(E545K),
PIK3CA(H1047L), PIK3CA(H1047Y), PIK3CA(M1043I), PIK3CA(Q546K),
PIK3CB, PIK3CD, PIK3CG, PIK4CB, PIM1, PIM2, PIM3, PIP5K1A, PIP5K2B,
PKAC-ALPHA, PKAC-BETA, PKMYT1, PKN1, PKN2, PLK1, PLK2, PLK3, PLK4,
PRKCD, PRKCE, PRKCH, PRKCQ, PRKD1, PRKD3, PRKG1, PRKG2, PRKR, PRKX,
PRP4, PYK2, QSK, RAF1, RET, RET(M918T), RET(V804L), RET(V804M),
RIOK1, RIOK2, RIOK3, RIPK1, RIPK2, RIPK4, ROCK1, ROCK2, ROS1,
RPS6KA1(Kin.Dom.1-N-terminal), RPS6KA1(Kin.Dom.2-C-terminal),
RPS6KA2(Kin.Dom.1-N-terminal), RPS6KA2(Kin.Dom.2-C-terminal),
RPS6KA3(Kin.Dom.1-N-terminal), RPS6KA4(Kin.Dom.1-N-terminal),
RPS6KA4(Kin.Dom.2-C-terminal), RPS6KA5(Kin.Dom.1-N-terminal),
RPS6KA5(Kin.Dom.2-C-terminal), RPS6KA6(Kin.Dom.1-N-terminal),
RPS6KA6(Kin.Dom.2-C-terminal), SBK1, SgK085, SgK110, SIK, SIK2,
SLK, SNARK, SRC, SRMS, SRPK1, SRPK2, SRPK3, STK16, STK33, STK39,
SYK, TAK1, TAO1, TAOK2, TAOK3, TBK1, TEC, TESK1, TGFBR1, TGFBR2,
TIE1, TIE2, TLK1, TLK2, TNIK, TNK1, TNK2, TNNI3K, TRKA, TRKB, TRKC,
TSSK1B, TTK, TXK, TYK2(JH1domain-catalytic),
TYK2(JH2domain-pseudokinase), TYRO3, ULK1, ULK2, ULK3, VEGFR2,
WEE1, WEE2, YANK2, YANK3, YES, YSK1, YSK4, ZAK and ZAP70. Compounds
of the invention are screened against the kinase panel (wild type
and/or mutation thereof) and inhibit the activity of at least one
of said panel members.
Mutant forms of a kinase means single or multiple amino acid
changes from the wild-type sequence.
The term "subject" as used herein refers to a mammal. A subject
therefore refers to, for example, dogs, cats, horses, cows, pigs,
guinea pigs, and the like. Preferably the subject is a human. When
the subject is a human, the subject may be referred to herein as a
patient.
"Treat", "treating" and "treatment" refer to a method of
alleviating or abating a disease and/or its attendant symptoms.
As used herein, the term "pharmaceutically acceptable salt" refers
to those salts of the compounds formed by the process of the
present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. Pharmaceutically acceptable salts are well
known in the art. For example, S. M. Berge, et al. describes
pharmaceutically acceptable salts in detail in J. Pharmaceutical
Sciences, 66: 1-19 (1977). The salts can be prepared in situ during
the final isolation and purification of the compounds of the
invention, or separately by reacting the free base function with a
suitable organic acid. Examples of pharmaceutically acceptable
include, but are not limited to, nontoxic acid addition salts are
salts of an amino group formed with inorganic acids such as
hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid
and perchloric acid or with organic acids such as acetic acid,
maleic acid, tartaric acid, citric acid, succinic acid or malonic
acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include, but are
not limited to, adipate, alginate, ascorbate, aspartate,
benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like. Further
pharmaceutically acceptable salts include, when appropriate,
nontoxic ammonium, quaternary ammonium, and amine cations formed
using counterions such as halide, hydroxide, carboxylate, sulfate,
phosphate, nitrate, alkyl having from 1 to 6 carbon atoms,
sulfonate and aryl sulfonate.
As used herein, the term "pharmaceutically acceptable ester" refers
to esters of the compounds formed by the process of the present
invention which hydrolyze in vivo and include those that break down
readily in the human body to leave the parent compound or a salt
thereof. Suitable ester groups include, for example, those derived
from pharmaceutically acceptable aliphatic carboxylic acids,
particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic
acids, in which each alkyl or alkenyl moiety advantageously has not
more than 6 carbon atoms. Examples of particular esters include,
but are not limited to, formates, acetates, propionates, butyrates,
acrylates and ethylsuccinates.
The term "pharmaceutically acceptable prodrugs" as used herein
refers to those prodrugs of the compounds formed by the process of
the present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals with undue toxicity, irritation, allergic
response, and the like, commensurate with a reasonable benefit/risk
ratio, and effective for their intended use, as well as the
zwitterionic forms, where possible, of the compounds of the present
invention. "Prodrug", as used herein means a compound which is
convertible in vivo by metabolic means (e.g. by hydrolysis) to
afford any compound delineated by the formulae of the instant
invention. Various forms of prodrugs are known in the art, for
example, as discussed in Bundgaard, (ed.), Design of Prodrugs,
Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, vol.
4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed). "Design
and Application of Prodrugs, Textbook of Drug Design and
Development, Chapter 5, 113-191 (1991); Bundgaard, et al., Journal
of Drug Deliver Reviews, 8:1-38(1992); Bundgaard, J. of
Pharmaceutical Sciences, 77:285 et seq. (1988); Higuchi and Stella
(eds.) Prodrugs as Novel Drug Delivery Systems, American Chemical
Society (1975); and Bernard Testa & Joachim Mayer, "Hydrolysis
In Drug And Prodrug Metabolism: Chemistry, Biochemistry And
Enzymology," John Wiley and Sons, Ltd. (2002).
This invention also encompasses pharmaceutical compositions
containing, and methods of treating disorders through
administering, pharmaceutically acceptable prodrugs of compounds of
the invention. For example, compounds of the invention having free
amino, amido, hydroxy or carboxylic groups can be converted into
prodrugs. Prodrugs include compounds wherein an amino acid residue,
or a polypeptide chain of two or more (e.g., two, three or four)
amino acid residues is covalently joined through an amide or ester
bond to a free amino, hydroxy or carboxylic acid group of compounds
of the invention. The amino acid residues include but are not
limited to the 20 naturally occurring amino acids commonly
designated by three letter symbols and also includes
4-hydroxyproline, hydroxyysine, demosine, isodemosine,
3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,
citrulline, homocysteine, homoserine, ornithine and methionine
sulfone. Additional types of prodrugs are also encompassed. For
instance, free carboxyl groups can be derivatized as amides or
alkyl esters. Free hydroxy groups may be derivatized using groups
including but not limited to hemisuccinates, phosphate esters,
dimethylaminoacetates, and phosphoryloxymethyloxy carbonyls, as
outlined in Advanced Drug Delivery Reviews, 1996, 19, 1 15.
Carbamate prodrugs of hydroxy and amino groups are also included,
as are carbonate prodrugs, sulfonate esters and sulfate esters of
hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl
and (acyloxy)ethyl ethers wherein the acyl group may be an alkyl
ester, optionally substituted with groups including but not limited
to ether, amine and carboxylic acid functionalities, or where the
acyl group is an amino acid ester as described above, are also
encompassed. Prodrugs of this type are described in J. Med. Chem.
1996, 39, 10. Free amines can also be derivatized as amides,
sulfonamides or phosphonamides. All of these prodrug moieties may
incorporate groups including but not limited to ether, amine and
carboxylic acid functionalities
Combinations of substituents and variables envisioned by this
invention are only those that result in the formation of stable
compounds. The term "stable", as used herein, refers to compounds
which possess stability sufficient to allow manufacture and which
maintains the integrity of the compound for a sufficient period of
time to be useful for the purposes detailed herein (e.g.,
therapeutic or prophylactic administration to a subject).
Doublecortin-Like Kinases
Described herein are compounds that can inhibit at least one of
doublecortin-like kinases (DCLK), e.g., isoforms such as DCLK1 and
DCLK2, which are members of serine/threonine-protein kinase family
and Ca.sup.2+/calmodulin-dependent protein kinase class of
enzymes.
These doublecortin-like kinases (DCLK) commonly contain i) two
N-terminal doublecortin domains, which can bind or associate
microtubules; ii) a C-terminal serine/threonine protein kinase
domain, which shows substantial homology to
Ca.sup.2+/calmodulin-dependent protein kinase, and iii) a
serine/proline-rich domain in between the doublecortin and the
protein kinase domains, which mediates multiple protein-protein
interactions.
Human serine/threonine-protein kinase doublecortin-like kinase 1
(DCLK1) refers to the protein products of the human gene DCLK1.
This includes any serine/threonine-protein kinases (EC:2.7.11.1)
referred to as, for example, doublecortin and CaM kinase-like
1(DCAMKL1), doublecortin domain-containing protein 3A(DCDC.sub.3A),
CPG16 (candidate plasticity gene 16), CaMK-like CREB regulatory
kinase 1 (CL1, CLICK-I, CLICK1, or CLIK1) and/or KIAA0369, proteins
listed under UNIPROT ID: Q5VZY9 or uniprot ID: O15075, or
homologous proteins or isoforms thereof, including but not limited
to isoforms 1-4 (alternate nomenclature DCLK1-long,
DCLK1-short).
Likewise, human serine/threonine-protein kinase doublecortin-like
kinase 2 (DCLK2) refers to the protein products of the human gene
DCLK2. This includes any serine/threonine-protein kinases
(EC:2.7.11.1) which may be alternately named as, for example,
CaMK-like CREB regulatory kinase 2 (CL2, CLICK-II, CLICK2, or
CLIK2); doublecortin and CaM kinase-like 2 (DCAMKL2); doublecortin
domain-containing protein 3B (DCDC.sub.3B); doublecortin-like and
CAM kinase-like 2 (DCLK2); doublecortin-like kinase 2 (DCK2);
serine/threonine-protein kinase DCLK2 and proteins listed under
UNIPROT ID: Q8N568, or homologous proteins or isoforms thereof.
Isoforms include but are not limited to isoforms 1-3 listed in
UNIPROT.
DCLK1 is a microtubule-associated protein that plays a key role in
neuronal migration, retrograde transport, neuronal apoptosis,
neurogenesis and synapse maturation by regulation of mitotic
spindle formation, which can be independent from its protein kinase
activity (Reiner, O. et al, BMC Genomics 2006, 7 (1), 1-16).
Moreover, DCLK1 is frequently mutated across many cancer types. In
particular, DCLK1 can be a driver gene for gastic cancer and is
listed among the top 15 presumed driver genes (The Cancer Genome
Atlas Research, Nature 2014, 513 (7517), 202-209). Expression of
DCLK1 is also associated with poor prognosis in gastric cancer
patients (Gy rffy, B. et al., PLoS ONE 2013, 8 (12)). Additionally,
DCLK1 can be a marker of quiescent stem-cells in the stomach,
intestine and pancreas, and those cells have been shown to function
as the cancer stem cells which initiate and support cancers in the
intestine and pancreas, alone, or when harboring oncogenic
mutations, or upon loss of tumour suppressor genes. For instance,
ablation of these cells induces rapid regression of tumors in
murine models of intestinal cancer (Ito, H. et al., PLoS ONE 2016,
11 (1); Nakanishi, Y. et al., Nature genetics 2013, 45 (1), 98-103;
Westphalen, C. B., et. al, The Journal of clinical investigation
2014, 124 (3), 1283-95; Westphalen, C. B. et al., Cell Stem Cell 18
(4), 441-455).
Accordingly, as described herein, DCLK1/2 positive cells and
DCLK1/2 functions can be targeted as promising therapeutic
strategies in cancer treatments (e.g. treatments for gastric,
pancreatic and intestinal cancers).
However, currently available DCLK1/2 kinase inhibitors are highly
multi-targeted and have been shown to inhibit a number of other
kinases and bromodomains. Prior to the invention described herein,
there were no potent or selective inhibitors of the DCLK1/2 kinases
described in the literature.
Accordingly, described herein are series of compounds based around
a pyrimido-diazepinone scaffold. Such compounds, including those
based on a
2-amino-5,8,11-trimethyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diaz-
epin-6-one scaffold, can be potent and selective inhibitors of
DCLK1 or DCLK2, or alternatively, selective dual inhibitors of
DCLK1 and DCLK2, thereby providing a method of treating/preventing
a related disease mediated by DCLK1 and/or DCLK2.
DCLK1/2 Inhibitor Compounds
Described herein are series of compounds based around a
pyrimido-diazepinone scaffold. Such compounds can be inhibitors of
kinases (e.g., DCLK, including DCLK isoforms such as DCLK1 and/or
DCLK2) and are referred to herein as "DCLK1/2 inhibitor
compounds."
In embodiments, the invention provides a compound of formula
F-1:
##STR00031##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.5 is hydrogen, optionally substituted alkyl, optionally
substituted aralkyl, or optionally substituted carbocyclic; and
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In embodiments, R.sub.5 is methyl.
In embodiments, R.sub.2 is unsubstituted alkyl.
In certain embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen.
In certain embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In embodiments, p is 0; or when p is 1, R.sub.7 is unsubstituted
alkyl.
In certain embodiments, R.sub.1 is methyl, ethyl, propyl,
iso-propyl, butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl,
piperidinyl, pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl,
pyrimidinyl, pyrazinyl, pyridizinyl, quinolinyl, thienyl,
thiazolyl, oxazolyl, isoxazolyl, pyrrolyl, furanyl, isoquinolinyl,
imiazolyl, or triazolyl, each of which may be optionally
substituted.
In a further embodiment, R.sub.1 is phenyl or pyridyl, each of
which may be optionally substituted.
In another embodiment, R.sub.1 is substituted with 0-4
substituents, selected from halo, nitro, cyano, hydroxyl, amino,
NH(R.sub.A), N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A,
C(O)OR.sub.A, C(O)NH.sub.2, C(O)NH(R.sub.A),
C(O)N(R.sub.A)(R.sub.A), alkyl, aryl, arylalkyl, alkoxy,
heteroaryl, heterocyclic, and carbocyclic, each of which may be
further substituted; wherein each R.sub.A is independently selected
from alkyl, alkenyl, carbocyclic, aryl, heteroaryl, and
heterocyclic, or two R.sub.A on the same atom combine to form a
heterocyclic, each of which may be further substituted.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy, CO.sub.2Me,
##STR00032##
In embodiments, R.sub.1 is phenyl, pyridyl, pyrimidinyl, furyl,
pyrrolyl, pyrazolyl, imidazolyl, thienyl, or
bicyclo[1.1.1]pent-1-yl, each of which may be optionally
substituted.
In embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, carbocyclic,
SO.sub.2(R.sub.A), SO.sub.3(R.sub.A), SO.sub.2N(R.sub.A)(R.sub.A),
SO.sub.2NH(R.sub.A), SO.sub.2NH.sub.2, PO(OR.sub.A)(OR.sub.A), or
PO(OR.sub.A)(R.sub.A), each of which may be further substituted,
and wherein each R.sub.A is independently selected from alkyl,
alkenyl, carbocyclic, aryl, heteroaryl, and heterocyclic, or two
R.sub.A on the same atom combine to form a heterocyclic, each of
which may be further substituted.
In certain embodiments, R.sub.1 is selected from the group
consisting of
##STR00033##
In embodiments, the compound is of formula F-1-a:
##STR00034##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted.
In embodiments, R.sub.2 is unsubstituted alkyl.
In certain embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen.
In certain embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In certain embodiments, R.sub.1 is methyl, ethyl, propyl,
iso-propyl, butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl,
piperidinyl, pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl,
pyrimidinyl, pyrazinyl, pyridizinyl, quinolinyl, thienyl,
thiazolyl, oxazolyl, isoxazolyl, pyrrolyl, furanyl, isoquinolinyl,
imiazolyl, or triazolyl, each of which may be optionally
substituted.
In a further embodiment, R.sub.1 is phenyl or pyridyl, each of
which may be optionally substituted.
In another embodiment, R.sub.1 is substituted with 0-4
substituents, selected from halo, nitro, cyano, hydroxyl, amino,
NH(R.sub.A), N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A,
C(O)OR.sub.A, C(O)NH.sub.2, C(O)NH(R.sub.A),
C(O)N(R.sub.A)(R.sub.A), alkyl, aryl, arylalkyl, alkoxy,
heteroaryl, heterocyclic, and carbocyclic, each of which may be
further substituted; wherein each R.sub.A is independently selected
from alkyl, alkenyl, carbocyclic, aryl, heteroaryl, and
heterocyclic, or two R.sub.A on the same atom combine to form a
heterocyclic, each of which may be further substituted.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy, CO.sub.2Me,
##STR00035##
In embodiments, R.sub.1 is phenyl, pyridyl, pyrimidinyl, furyl,
pyrrolyl, pyrazolyl, imidazolyl, thienyl, or
bicyclo[1.1.1]pent-1-yl, each of which may be optionally
substituted.
In embodiments, R.sub.1 is substituted with 0-4 substituents,
selected from halo, nitro, cyano, hydroxyl, amino, NH(R.sub.A),
N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A, C(O)OR.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, carbocyclic,
SO.sub.2(R.sub.A), SO.sub.3(R.sub.A), SO.sub.2N(R.sub.A) (R.sub.A),
SO.sub.2NH(R.sub.A), SO.sub.2NH.sub.2, PO(OR.sub.A)(OR.sub.A), or
PO(OR.sub.A)(R.sub.A), each of which may be further substituted,
and wherein each R.sub.A is independently selected from alkyl,
alkenyl, carbocyclic, aryl, heteroaryl, and heterocyclic, or two
R.sub.A on the same atom combine to form a heterocyclic, each of
which may be further substituted.
In embodiments, R.sub.1 is selected from the group consisting
of
##STR00036##
In embodiments, the compound is of formula F-1-b:
##STR00037##
or a pharmaceutically acceptable salt, ester or prodrug
thereof.
In embodiments, R.sub.2 is unsubstituted alkyl.
In certain embodiments, R.sub.2 is methyl, ethyl, propyl, or
iso-propyl, each of which may be optionally substituted with one or
more halogen such as F.
In certain embodiments, R.sub.2 is --CH.sub.2--CH.sub.2F,
--CH.sub.2--CHF.sub.2, or --CH.sub.2--CF.sub.3.
In certain embodiments, R' is selected from alkoxy, CO.sub.2Me,
##STR00038##
In certain embodiments, q is 1 or 2.
In certain embodiments, when q is 2, one R' is --OCH.sub.3.
In embodiments, the compound has the structure of any one of
Compounds 1-21 as described herein, or a pharmaceutically
acceptable salt, ester or prodrug thereof.
In embodiments, the compound has a structure selected from the
group consisting of:
##STR00039## ##STR00040## ##STR00041## ##STR00042## ##STR00043##
##STR00044## or a pharmaceutically acceptable salt, ester or
prodrug thereof.
In embodiments, the compound has a structure according to formula
A-1:
##STR00045##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
X is CHR.sub.4, CR.sub.4, NH, NR.sub.4 or N;
Y is NR.sub.5, N, S, SO, SO.sub.2, O, CHR.sub.5, or CR.sub.5;
wherein at least one of X and Y is NH, NR.sub.4, NR.sub.5, N, S,
SO, SO.sub.2, or O;
A is a single bond or double bond;
B is a single bond or double bond, wherein both A and B are not
double bonds;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.4 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.5 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
or R.sub.3 and X, together with the atoms to which they are
attached, form a 3-8 membered carbocyclic, aryl, heterocyclic, or
heteroaryl; each of which is optionally substituted;
or X and Y, together with the atoms to which they are attached,
form a 3-8 membered carbocyclic, aryl, heterocyclic, or heteroaryl;
each of which is optionally substituted; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In certain embodiments, the invention provides a compound wherein X
is CR.sub.4 or CHR.sub.4, and Y is NR.sub.5.
In other embodiments, the invention provides a compound wherein
R.sub.4 is hydrogen, alkyl, aryl, heteroaryl, heterocyclic, or
carbocyclic, each of which may be optionally substituted; and
R.sub.5 is hydrogen, alkyl, aryl, heteroaryl, heterocyclic, or
carbocyclic, each of which may be optionally substituted.
In certain embodiments, the invention provides a compound wherein X
and Y, together with the atoms to which they are attached, form a
3-8 membered cycloalkyl, aryl, heterocycloalkyl, or heteroaryl;
each of which is optionally substituted.
In other embodiments, the invention provides a compound wherein
R.sub.3 and X, together with the atoms to which they are attached,
form a 3-8 membered cycloalkyl, aryl, heterocycloalkyl, or
heteroaryl; each of which is optionally substituted.
In some embodiments, the invention provides a compound wherein X is
N and Y is CR.sub.5.
In a further embodiment, R.sub.5 is alkyl, aryl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted.
In embodiments, the invention provides a compound of B-1:
##STR00046##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is aryl, or heteroaryl, wherein R.sub.1 may be optionally
substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen or methyl;
R.sub.4 is hydrogen or methyl; and
R.sub.6 is hydrogen.
In one embodiment, R.sub.1 is phenyl or pyridyl, each of which may
be optionally substituted.
In a further embodiment, R.sub.1 is substituted with 0-4
substituents, selected from N(R.sub.A)(R.sub.A), C(O)NH(R.sub.A),
alkoxy, and heterocyclic, each of which may be further substituted;
wherein each R.sub.A is independently selected from alkyl, and
heterocyclic. In another further embodiment, R.sub.1 is substituted
with 0-4 substituents, selected from alkoxy,
##STR00047##
In embodiments, the invention provides a compound of formula
C-1:
##STR00048##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is aryl, heteroaryl, which may be optionally
substituted;
R.sub.2 is hydrogen or methyl;
R.sub.3 is hydrogen;
R.sub.4 is hydrogen; and
R.sub.6 is hydrogen.
In certain embodiments, R.sub.1 is phenyl or pyridyl, each of which
may be optionally substituted.
In a further embodiment, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy, or heterocyclic, which may be
further substituted.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy,
##STR00049##
In embodiments, the invention provides a compound of formula
D-1:
##STR00050##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-6.
In one embodiment, R.sub.1 is methyl, ethyl, propyl, iso-propyl,
butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl, piperidinyl,
pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridizinyl, quinolinyl, thienyl, thiazolyl, oxazolyl,
isoxazolyl, pyrrolyl, furanyl, isoquinolinyl, imiazolyl, or
triazolyl, each of which may be optionally substituted.
In a further embodiment, R.sub.1 is alkyl, phenyl, cyclohexyl,
piperidinyl, quinolinyl, or pyridyl, each of which may be
optionally substituted.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from halo, nitro, cyano, hydroxyl, amino,
NH(R.sub.A), N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A,
C(O)OR.sub.A, C(O)NH.sub.2, C(O)NH(R.sub.A),
C(O)N(R.sub.A)(R.sub.A), alkyl, aryl, arylalkyl, alkoxy,
heteroaryl, heterocyclic, and carbocyclic, each of which may be
further substituted; wherein each R.sub.A is independently selected
from alkyl, carbocyclic, aryl, heteroaryl, and heterocyclic.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from alkyl, alkoxy, hydroxyl,
##STR00051##
In embodiments, the invention provides a compound of formula
E-1:
##STR00052##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen or optionally substituted alkyl;
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-6.
In certain embodiments, R.sub.1 is methyl, ethyl, propyl,
iso-propyl, butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl,
piperidinyl, pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl,
pyrimidinyl, pyrazinyl, pyridizinyl, quinolinyl, thienyl,
thiazolyl, oxazolyl, isoxazolyl, pyrrolyl, furanyl, isoquinolinyl,
imiazolyl, or triazolyl, each of which may be optionally
substituted.
In a further embodiment, R.sub.1 is phenyl or pyridyl, each of
which may be optionally substituted.
In another further embodiment, R.sub.1 is substituted with 0-4
substituents, selected from halo, nitro, cyano, hydroxyl, amino,
NH(R.sub.A), N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A,
C(O)OR.sub.A, C(O)NH.sub.2, C(O)NH(R.sub.A),
C(O)N(R.sub.A)(R.sub.A), alkyl, aryl, arylalkyl, alkoxy,
heteroaryl, heterocyclic, and carbocyclic, each of which may be
further substituted; wherein each R.sub.A is independently selected
from alkyl, carbocyclic, aryl, heteroaryl, and heterocyclic.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy,
##STR00053##
In embodiments, the invention provides a compound of formula
F-I-1:
##STR00054##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
Y is S, SO, SO.sub.2, or O;
R.sub.1 is alkyl, aryl, heteroaryl, heterocyclic, or carbocyclic,
wherein R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.6 is hydrogen or optionally substituted alkyl;
each R.sub.7 is independently alkyl, alkenyl, aryl, arylalkyl,
heteroaryl, heterocyclic, carbocyclic, alkoxy, NH(alkyl), NH(aryl),
N(alkyl)(alkyl), or N(alkyl)(aryl), each of which may be optionally
substituted; halo, nitro, or cyano; and
p is 0-4.
In one embodiment, R.sub.1 is methyl, ethyl, propyl, iso-propyl,
butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl, piperidinyl,
pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridizinyl, quinolinyl, thienyl, thiazolyl, oxazolyl,
isoxazolyl, pyrrolyl, furanyl, isoquinolinyl, imiazolyl, or
triazolyl, each of which may be optionally substituted.
In a further embodiment, R.sub.1 is phenyl or pyridyl, each of
which may be optionally substituted.
In another embodiment, R.sub.1 is substituted with 0-4
substituents, selected from halo, nitro, cyano, hydroxyl, amino,
NH(R.sub.A), N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A,
C(O)OR.sub.A, C(O)NH.sub.2, C(O)NH(R.sub.A),
C(O)N(R.sub.A)(R.sub.A), alkyl, aryl, arylalkyl, alkoxy,
heteroaryl, heterocyclic, and carbocyclic, each of which may be
further substituted;
wherein each R.sub.A is independently selected from alkyl,
carbocyclic, aryl, heteroaryl, and heterocyclic.
In a further embodiment, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy, CO.sub.2Me,
##STR00055##
In embodiments, the invention provides a compound of formula
G-1:
##STR00056##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.1 is alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or 3
heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic, wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted;
R.sub.5 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heterocyclic, or carbocyclic, each of which may be optionally
substituted; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In one embodiment, R.sub.1 is methyl, ethyl, propyl, iso-propyl,
butyl, s-butyl, t-butyl, pentyl, hexyl, cyclohexyl, piperidinyl,
pyrrolidino, phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridizinyl, quinolinyl, thienyl, thiazolyl, oxazolyl,
isoxazolyl, pyrrolyl, furanyl, isoquinolinyl, imiazolyl, or
triazolyl, each of which may be optionally substituted.
In a further embodiment, R.sub.1 is optionally substituted
phenyl.
In another embodiment, R.sub.1 is substituted with 0-4
substituents, selected from halo, nitro, cyano, hydroxyl, amino,
NH(R.sub.A), N(R.sub.A)(R.sub.A), CO.sub.2H, C(O)R.sub.A,
C(O)NH.sub.2, C(O)NH(R.sub.A), C(O)N(R.sub.A)(R.sub.A), alkyl,
aryl, arylalkyl, alkoxy, heteroaryl, heterocyclic, and carbocyclic,
each of which may be further substituted; wherein each R.sub.A is
independently selected from alkyl, carbocyclic, aryl, heteroaryl,
and heterocyclic.
In certain embodiments, R.sub.1 is substituted with 0-4
substituents, selected from alkoxy, hydroxyl,
##STR00057##
In another embodiment, R.sub.5 is optionally substituted phenyl or
optionally substituted cyclopentyl.
In embodiments, the invention provides a compound of formula
I-2:
##STR00058##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
A is a single bond or double bond;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent (for example, halogen, --OH,
--NO.sub.2, --CN, --NH.sub.2, protected amino,
--NH--C.sub.1-C.sub.12-alkyl, --NH--C.sub.2-C.sub.12-alkenyl,
--NH--C.sub.2-C.sub.12-alkenyl, --NH--C.sub.3-C.sub.12-cycloalkyl,
--NH-aryl, --NH-heteroaryl, --NH-heterocycloalkyl, -dialkylamino,
-diarylamino, -diheteroarylamino, --O--C.sub.1-C.sub.12-alkyl,
--O-C.sub.2-C.sub.12-alkenyl, --O--C.sub.2-C.sub.12-alkenyl,
--O--C.sub.3-C.sub.12-cycloalkyl, --O-aryl, --O-heteroaryl,
--O-heterocycloalkyl, --C(O)--C.sub.1-C.sub.12-alkyl,
--C(O)--C.sub.2-C.sub.12-alkenyl, --C(O)--C.sub.2-C.sub.12-alkenyl,
--C(O)--C.sub.3-C.sub.12-cycloalkyl, --C(O)-aryl,
--C(O)-heteroaryl, --C(O)-heterocycloalkyl, --CONH.sub.2,
--CONH--C.sub.1-C.sub.12-alkyl, --CONH--C.sub.2-C.sub.12-alkenyl,
--CONH--C.sub.2-C.sub.12-alkenyl,
--CONH--C.sub.3-C.sub.12-cycloalkyl, --CONH-aryl,
--CONH-heteroaryl, --CONH-heterocycloalkyl, --OCO.sub.2--
C.sub.1-C.sub.12-alkyl, --OCO.sub.2--C.sub.2-C.sub.12-alkenyl,
--OCO.sub.2--C.sub.2-C.sub.12-alkenyl,
--OCO2-C.sub.3-C.sub.12-cycloalkyl, --OCO.sub.2-aryl,
--OCO.sub.2-heteroaryl, --OCO.sub.2-heterocycloalkyl,
--OCONH.sub.2, --OCONH--C.sub.1-C.sub.12-alkyl,
OCONH--C.sub.2-C.sub.12-alkenyl, --OCONH--C.sub.2-C.sub.12-alkenyl,
--OCONH--C.sub.3-C.sub.12-cycloalkyl, --OCONH-aryl,
--OCONH-heteroaryl, --OCONH-heterocycloalkyl,
--NHC(O)--C.sub.1-C.sub.12-alkyl,
--NHC(O)--C.sub.2-C.sub.12-alkenyl,
--NHC(O)--C.sub.2-C.sub.12-alkenyl,
--NHC(O)--C.sub.3-C.sub.12-cycloalkyl, --NHC(O)-aryl,
--NHC(O)-heteroaryl, --NHC(O)-heterocycloalkyl,
--NHCO.sub.2--C.sub.1-C.sub.12-alkyl,
--NHCO.sub.2--C.sub.2-C.sub.12-alkenyl, --NHCO.sub.2--
C.sub.2-C.sub.12-alkenyl,
--NHCO.sub.2--C.sub.3-C.sub.12-cycloalkyl, --NHCO.sub.2-aryl,
--NHCO.sub.2-heteroaryl, --NHCO.sub.2-heterocycloalkyl,
--NHC(O)NH.sub.2, --NHC(O)NH--C.sub.1-C.sub.12-alkyl,
--NHC(O)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(O)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(O)NH--C.sub.3-C.sub.12-cycloalkyl, --NHC(O)NH-aryl,
--NHC(O)NH-heteroaryl, --NHC(O)NH-heterocycloalkyl, NHC(S)NH.sub.2,
--NHC(S)NH-C.sub.1-C.sub.12-alkyl,
--NHC(S)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(S)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(S)NH--C.sub.3-C.sub.12-cycloalkyl, --NHC(S)NH-aryl,
--NHC(S)NH-heteroaryl, --NHC(S)NH-heterocycloalkyl,
--NHC(NH)NH.sub.2, --NHC(NH)NH--C.sub.1-C.sub.12-alkyl,
--NHC(NH)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)NH--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)NH--C.sub.3-C.sub.12-cycloalkyl, --NHC(NH)NH-aryl,
--NHC(NH)NH-heteroaryl, --NHC(NH)NH-- heterocycloalkyl,
--NHC(NH)--C.sub.1-C.sub.12-alkyl,
--NHC(NH)--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)--C.sub.2-C.sub.12-alkenyl,
--NHC(NH)--C.sub.3-C.sub.12-cycloalkyl, --NHC(NH)-aryl, --NHC(NH)--
heteroaryl, --NHC(NH)-heterocycloalkyl,
--C(NH)NH--C.sub.1-C.sub.12-alkyl,
--C(NH)NH-C.sub.2-C.sub.12-alkenyl,
--C(NH)NH--C.sub.2-C.sub.12-alkenyl,
--C(NH)NH--C.sub.3-C.sub.12-cycloalkyl, --C(NH)NH-aryl,
--C(NH)NH-heteroaryl, --C(NH)NH-heterocycloalkyl,
--S(O)--C.sub.1-C.sub.12-alkyl, --S(O)--C.sub.2-C.sub.12-alkenyl,
--S(O)--C.sub.2-C.sub.12-alkenyl,
--S(O)--C.sub.3-C.sub.12-cycloalkyl, --S(O)-aryl, --S(O)--
heteroaryl, --S(O)-heterocycloalkyl --SO.sub.2NH.sub.2,
--SO.sub.2NH--C.sub.1-C.sub.12-alkyl,
--SO.sub.2NH--C.sub.2-C.sub.12-alkenyl,
--SO.sub.2NH--C.sub.2-C.sub.12-alkenyl,
--SO.sub.2NH--C.sub.3-C.sub.12-cycloalkyl, --SO.sub.2NH-aryl,
--SO.sub.2NH-heteroaryl, --SO.sub.2NH-heterocycloalkyl,
--NHSO.sub.2--C.sub.1-C.sub.12-alkyl,
--NHSO.sub.2--C.sub.2-C.sub.12-alkenyl,
--NHSO.sub.2--C.sub.2-C.sub.12-alkenyl,
--NHSO.sub.2--C.sub.3-C.sub.12-cycloalkyl, --NHSO.sub.2-aryl,
--NHSO.sub.2-heteroaryl, --NHSO.sub.2-heterocycloalkyl,
--CH.sub.2NH.sub.2, --CH.sub.2SO.sub.2CH.sub.3, aryl, arylalkyl,
heteroaryl, heteroarylalkyl, heterocycloalkyl,
--C.sub.3-C.sub.12-cycloalkyl, polyalkoxyalkyl, polyalkoxy,
-methoxymethoxy, -methoxyethoxy, --SH, --S--C.sub.1-C.sub.12-alkyl,
--S--C.sub.2-C.sub.12-alkenyl, --S--C.sub.2-C.sub.12-alkenyl,
--S--C.sub.3-C.sub.12-cycloalkyl, --S-aryl, --S-- heteroaryl,
--S-heterocycloalkyl, or methylthiomethyl);
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen, optionally substituted alkyl (including
aralkyl), optionally substituted cycloalkyl, and optionally
substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of formula
II-2:
##STR00059##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
E is NR.sub.2 or CHR.sub.2;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl (including aralkyl), optionally substituted
cycloalkyl, and optionally substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In certain embodiments, E is NR.sub.2. In certain embodiments,
R.sub.2 is H or --CH.sub.3.
In embodiments, the invention provides a compound of formula
III-2:
##STR00060##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl (including aralkyl), optionally substituted
cycloalkyl, and optionally substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of formula
IV-2:
##STR00061##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl (including aralkyl), optionally substituted
cycloalkyl, and optionally substituted heterocyclyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of formula
V-2:
##STR00062##
or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
R.sub.2 is hydrogen or optionally substituted alkyl;
R.sub.3 is --OH or --O-(optionally substituted alkyl);
R.sub.4 is hydrogen or optionally substituted alkyl; and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of Formula
VI-2:
##STR00063## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is, independently for each occurrence, hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl; or
two X moieties on adjacent atoms of the thiophene ring can form,
together with the atoms to which they are attached, a phenyl ring;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of Formula
VII-2:
##STR00064## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen, optionally substituted alkyl, optionally
substituted cycloalkyl, and optionally substituted heterocyclyl;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of Formula
VIII-2:
##STR00065## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
X is an optional substituent as defined for formula I;
Z is O or S;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 is hydrogen, optionally substituted alkyl, optionally
substituted cycloalkyl, and optionally substituted heterocyclyl;
and
R.sub.6 is hydrogen or optionally substituted alkyl.
In embodiments, the invention provides a compound of Formula
IX-2:
##STR00066## or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein,
A is a single bond or double bond;
R' is H or alkyl;
L is absent, S, SO, SO.sub.2, or CO;
Y is hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, and optionally substituted heterocyclyl;
R.sub.1 is H, alkyl, alkenyl, alkynyl, each containing 0, 1, 2, or
3 heteroatoms selected from O, S, or N; or R.sub.1 is aryl,
arylalkyl, heteroaryl, heterocyclic, or carbocyclic; wherein
R.sub.1 may be optionally substituted;
R.sub.2 and R.sub.2' are each independently hydrogen, optionally
substituted alkyl, optionally substituted cycloalkyl, and
optionally substituted heterocyclyl;
or Y and R.sub.2' can form, together with the atoms to which they
are attached, a five-membered ring; and
R.sub.6 is hydrogen or optionally substituted alkyl.
Exemplary methods for preparation or synthesizing of these
compounds are described herein and in, e.g., International
Publication Nos. WO2010/080712 and WO2014145909, each of which is
incorporated by reference in its entirety.
Another embodiment is a method of making a compound of any of the
formulae herein using any one, or combination of, reactions
delineated herein. The method can include the use of one or more
intermediates or chemical reagents delineated herein.
Another aspect is an isotopically labeled compound of any of the
formulae delineated herein. Such compounds have one or more isotope
atoms which may or may not be radioactive (e.g., .sup.3H, .sup.2H,
.sup.14C, .sup.13C, .sup.35S, .sup.32P, .sup.125I, and .sup.131I)
introduced into the compound. Such compounds are useful for drug
metabolism studies and diagnostics, as well as therapeutic
applications.
A compound of the invention can be prepared as a pharmaceutically
acceptable acid addition salt by reacting the free base form of the
compound with a pharmaceutically acceptable inorganic or organic
acid. Alternatively, a pharmaceutically acceptable base addition
salt of a compound of the invention can be prepared by reacting the
free acid form of the compound with a pharmaceutically acceptable
inorganic or organic base.
Alternatively, the salt forms of the compounds of the invention can
be prepared using salts of the starting materials or
intermediates.
The free acid or free base forms of the compounds of the invention
can be prepared from the corresponding base addition salt or acid
addition salt from, respectively. For example, a compound of the
invention in an acid addition salt form can be converted to the
corresponding free base by treating with a suitable base (e.g.,
ammonium hydroxide solution, sodium hydroxide, and the like). A
compound of the invention in a base addition salt form can be
converted to the corresponding free acid by treating with a
suitable acid (e.g., hydrochloric acid, etc.).
Prodrug derivatives of the compounds of the invention can be
prepared by methods known to those of ordinary skill in the art
(e.g., for further details see Saulnier et al., (1994), Bioorganic
and Medicinal Chemistry Letters, Vol. 4, p. 1985). For example,
appropriate prodrugs can be prepared by reacting a non-derivatized
compound of the invention with a suitable carbamylating agent
(e.g., 1,1-acyloxyalkylcarbanochloridate, para-nitrophenyl
carbonate, or the like).
Protected derivatives of the compounds of the invention can be made
by means known to those of ordinary skill in the art. A detailed
description of techniques applicable to the creation of protecting
groups and their removal can be found in T. W. Greene, "Protecting
Groups in Organic Chemistry", 3rd edition, John Wiley and Sons,
Inc., 1999.
Compounds of the present invention can be conveniently prepared, or
formed during the process of the invention, as solvates (e.g.,
hydrates). Hydrates of compounds of the present invention can be
conveniently prepared by recrystallization from an aqueous/organic
solvent mixture, using organic solvents such as dioxin,
tetrahydrofuran or methanol.
Acids and bases useful in the methods herein are known in the art.
Acid catalysts are any acidic chemical, which can be inorganic
(e.g., hydrochloric, sulfuric, nitric acids, aluminum trichloride)
or organic (e.g., camphorsulfonic acid, p-toluenesulfonic acid,
acetic acid, ytterbium triflate) in nature. Acids are useful in
either catalytic or stoichiometric amounts to facilitate chemical
reactions. Bases are any basic chemical, which can be inorganic
(e.g., sodium bicarbonate, potassium hydroxide) or organic (e.g.,
triethylamine, pyridine) in nature. Bases are useful in either
catalytic or stoichiometric amounts to facilitate chemical
reactions.
In addition, some of the compounds of this invention have one or
more double bonds, or one or more asymmetric centers. Such
compounds can occur as racemates, racemic mixtures, single
enantiomers, individual diastereomers, diastereomeric mixtures, and
cis- or trans- or E- or Z-double isomeric forms, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino
acids. All such isomeric forms of these compounds are expressly
included in the present invention. Optical isomers may be prepared
from their respective optically active precursors by the procedures
described above, or by resolving the racemic mixtures. The
resolution can be carried out in the presence of a resolving agent,
by chromatography or by repeated crystallization or by some
combination of these techniques which are known to those skilled in
the art. Further details regarding resolutions can be found in
Jacques, et al., Enantiomers, Racemates, and Resolutions (John
Wiley & Sons, 1981). The compounds of this invention may also
be represented in multiple tautomeric forms, in such instances, the
invention expressly includes all tautomeric forms of the compounds
described herein (e.g., alkylation of a ring system may result in
alkylation at multiple sites, the invention expressly includes all
such reaction products). When the compounds described herein
contain olefinic double bonds or other centers of geometric
asymmetry, and unless specified otherwise, it is intended that the
compounds include both E and Z geometric isomers. Likewise, all
tautomeric forms are also intended to be included. The
configuration of any carbon-carbon double bond appearing herein is
selected for convenience only and is not intended to designate a
particular configuration unless the text so states; thus a
carbon-carbon double bond depicted arbitrarily herein as trans may
be cis, trans, or a mixture of the two in any proportion. All such
isomeric forms of such compounds are expressly included in the
present invention. All crystal forms of the compounds described
herein are expressly included in the present invention.
The synthesized compounds can be separated from a reaction mixture
and further purified by a method such as column chromatography,
high pressure liquid chromatography, or recrystallization. As can
be appreciated by the skilled artisan, further methods of
synthesizing the compounds of the formulae herein will be evident
to those of ordinary skill in the art. Additionally, the various
synthetic steps may be performed in an alternate sequence or order
to give the desired compounds. In addition, the solvents,
temperatures, reaction durations, etc. delineated herein are for
purposes of illustration only and one of ordinary skill in the art
will recognize that variation of the reaction conditions can
produce the desired bridged macrocyclic products of the present
invention. Synthetic chemistry transformations and protecting group
methodologies (protection and deprotection) useful in synthesizing
the compounds described herein are known in the art and include,
for example, those such as described in R. Larock, Comprehensive
Organic Transformations, VCH Publishers (1989); T. W. Greene and
P.G.M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed., John
Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's
Reagents for Organic Synthesis, John Wiley and Sons (1994); and L.
Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John
Wiley and Sons (1995), and subsequent editions thereof.
The compounds of this invention may be modified by appending
various functionalities via any synthetic means delineated herein
to enhance selective biological properties. Such modifications are
known in the art and include those which increase biological
penetration into a given biological system (e.g., blood, lymphatic
system, central nervous system), increase oral availability,
increase solubility to allow administration by injection, alter
metabolism and alter rate of excretion.
The compounds of the invention are defined herein by their chemical
structures and/or chemical names. Where a compound is referred to
by both a chemical structure and a chemical name, and the chemical
structure and chemical name conflict, the chemical structure is
determinative of the compound's identity.
The recitation of a listing of chemical groups in any definition of
a variable herein includes definitions of that variable as any
single group or combination of listed groups. The recitation of an
embodiment for a variable herein includes that embodiment as any
single embodiment or in combination with any other embodiments or
portions thereof.
Methods
In one aspect, the invention provides a method of treating a
disease in a subject mediated by a kinase that is doublecortin-like
kinase (DCLK1/2) comprising administering to the subject a DCLK
inhibitor compound (e.g. compound of formula F-1, F-1-a, F-1-b of
A-1) as described herein, or a pharmaceutically acceptable salt,
ester or prodrug thereof. For instance, the DCLK inhibitor compound
may selectively inhibit DCLK1 and/or DCLK2, with inhibition
constant (Ki) for inhibiting DCLK1 and/or DCLK2 less than about 1
.mu.M, less than about 500 nM, less than about 100 nM, less than
about 50 nM, less than about 40 nM, less than about 30 nM, less
than about 20 nM, or preferably, less than about 15 nM.
In another aspect, the invention provides a method for reducing
doublecortin-like kinase (DCLK1/2)-dependent cell growth comprising
contacting a cell with a DCLK inhibitor compound as described
herein, or a pharmaceutically acceptable salt, ester or prodrug
thereof.
In another aspect, the invention provides a method of inhibiting a
doublecortin-like kinase (DCLK1/2) in a subject identified as in
need of such treatment, comprising administering a DCLK inhibitor
compound as described herein, or a pharmaceutically acceptable
salt, ester or prodrug thereof.
In embodiments, the invention provides a method of inhibiting a
disease, wherein the disease is mediated by DCLK1. In one
embodiment, the invention provides a method of inhibiting a
disease, wherein the disease is mediated by DCLK2. In one
embodiment, the invention provides a method of inhibiting a
disease, wherein the disease is mediated by DCLK1 and DCLK2.
In another embodiment, the invention provides a method of
inhibiting a disease, wherein the disease is cancer or a
proliferation disease.
In some embodiments, the cancer is lung cancer, colon cancer,
breast cancer, prostate cancer, liver cancer, brain cancer, kidney
cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer,
gastric cancer, pancreatic cancer, glioma, hepatocellular
carcinoma, papillary renal carcinoma, head and neck squamous cell
carcinoma, leukemia, lymphoma, myeloma, or a solid tumor.
In some embodiments, the cancer is a blood-borne cancer (e.g.,
chronic lymphocytic leukemia (CLL), follicular lymphoma (FL) or
indolent non-Hodgkin's lymphoma (iNHL)). In an embodiment, the
cancer is chronic lymphocytic leukemia (CLL), follicular lymphoma
(FL), or indolent non-Hodgkin's lymphoma (iNHL).
In some embodiments, the cancer is pertinent to gastic organs,
gastrointestinal tract, or digestive organs including stomach,
small intestine, large intestine, tongue, salivary glands,
pancreas, liver, and gallbladder.
In some embodiments, the disease is Barretts' esophagus, esophageal
cancer, salivary gland malignancies, colon and colorectal cancer,
intestinal cancer, gastric cancer, pancreatic cancer, skin cancer
or neuroblastoma.
In some embodiments, the invention includes a method of reducing
recurrence and/or relapse of the cancer or proliferation disease as
described herein.
In some embodiments, the invention includes a method of reducing
migration and/or metathesis of the cancer or proliferation disease
as described herein.
In some embodiments, the invention includes a method of treating,
reducing or preventing resistant cancer cells and/or cancer stem
cells in patients suffered from or diagnosed with the cancer or
proliferation disease as described herein.
In embodiments, the disease is a liver disease.
In some embodiments, the disease is a fatty liver disease,
non-alcoholic fatty liver disease (NAFLD), non-alcoholic
steatohepatitis (NASH), cirrhosis, fatty liver disease resulting
from hepatitis, fatty liver disease resulting from obesity, fatty
liver disease resulting from diabetes, fatty liver disease
resulting from insulin resistance, fatty liver disease resulting
from hypertriglyceridemia, Abetalipoproteinemia, glycogen storage
diseases, Wolmans disease, or acute fatty liver of pregnancy.
In embodiments, the disease is a neurodegenerative disease.
In some embodiments, the disease is Alzheimer's disease (AD),
Parkinson's disease (PD), Huntington's (HD) diseases, amyotrophic
lateral sclerosis (ALS), spinal muscular atrophy (SMA),
schizophrenia, attention-deficit/hyperactivity disorder (ADHD),
fetal alcohol syndrome and diabetic encephalopathy.
In another aspect, the invention provides a method of treating a
kinase mediated disorder in a subject comprising: administering to
the subject identified as in need thereof a kinase inhibitor
compound as described herein, or a pharmaceutically acceptable
salt, ester or prodrug thereof.
In embodiments, a compound described herein is an inhibitor of
DCLK1. In embodiments, a compound described herein is an inhibitor
of DCLK2. In embodiments, a compound described herein is a
selective inhibitor of DCLK1. In embodiments, a compound described
herein is a selective inhibitor of DCLK2. In embodiments, a
compound described herein is a dual inhibitor of DCLK1 and DCLK2.
In embodiments, a compound described herein is a selective dual
inhibitor of DCLK1 and DCLK2.
In embodiments, the subject is administered an additional
therapeutic agent. In some embodiments, an additional therapeutic
agent is an anti-inflammatory agent. In some embodiments, an
additional therapeutic agent is a chemotherapy agent. In some
embodiments, an additional therapeutic agent is a monoclonal
antibody. In some embodiment, an additional therapeutic agent is a
therapeutic agent for liver disease. In some embodiments, an
additional therapeutic agent is a therapeutic agent for
neurodegenerative disease.
In a further embodiment, the compound and the additional
therapeutic agent are administered simultaneously or
sequentially.
In another aspect, the invention provides a method for reducing
kinase-dependent cell growth comprising contacting a cell with a
kinase inhibitor compound as described herein.
In other aspects, the invention provides a method of inhibiting
kinase in a subject identified as in need of such treatment,
comprising administering a kinase inhibitor compound as described
herein.
In embodiments, the invention provides a method wherein the subject
is a human.
In other embodiments, the invention provides a method wherein the
kinase inhibitor has a Ki for inhibiting DCLK1 and/or DCLK2 less
than about 1 .mu.M, less than about 500 nM, less than about 100 nM,
less than about 50 nM, less than about 40 nM, less than about 30
nM, less than about 20 nM, or less than about 15 nM.
In one embodiment, the invention provides a method of synthesizing
a kinase inhibitor compound as described herein. For example, the
compounds (e.g. compounds of formulae F-1, F-1-a and F-1-b) can be
synthesized as described herein (e.g., Schemes 1-4 in
Examples).
Another aspect of this invention provides compounds or compositions
that are inhibitors of protein kinases (e.g., DCLK, including DCLK1
and/or DCLK2), and thus are useful for the treatment of the
diseases, disorders, and conditions, along with other uses
described herein. In certain embodiments, these compositions
optionally further comprise one or more additional therapeutic
agents.
As inhibitors of protein kinases (e.g., DCLK, including DCLK1
and/or DCLK2), the compounds and compositions of this invention are
particularly useful for treating or lessening the severity of a
disease, condition, or disorder where a protein kinase is
implicated in the disease, condition, or disorder.
In one aspect, the present invention provides a method for treating
or lessening the severity of a disease, condition, or disorder
where a protein kinase is implicated in the disease state. In
another aspect, the present invention provides a method for
treating or lessening the severity of a kinase disease, condition,
or disorder where inhibition of enzymatic activity is implicated in
the treatment of the disease. In another aspect, this invention
provides a method for treating or lessening the severity of a
disease, condition, or disorder with compounds that inhibit
enzymatic activity by binding to the protein kinase. Another aspect
provides a method for treating or lessening the severity of a
kinase disease, condition, or disorder by inhibiting enzymatic
activity of the kinase with a protein kinase inhibitor.
In some embodiments, said method is used to treat or prevent a
condition selected from autoimmune diseases, inflammatory diseases,
cancers, tumors, malignant tumors, proliferative and
hyperproliferative diseases, immunologically-mediated diseases,
bone diseases, metabolic diseases, neurological and
neurodegenerative diseases, cardiovascular diseases, liver disease,
hormone related diseases, allergies, asthma, and neurodegenerative
diseases including Alzheimer's disease. In other embodiments, said
condition is selected from a proliferative disorder, liver disease
and a neurodegenerative disorder.
One aspect of this invention provides compounds that are useful for
the treatment of diseases, disorders, and conditions characterized
by excessive or abnormal cell proliferation. Such diseases include,
a proliferative or hyperproliferative disease, cancer, liver
disease and a neurodegenerative disease. Examples of proliferative
and hyperproliferative diseases include, without limitation,
cancer.
The term "cancer" includes, but is not limited to, the following
cancers: breast; ovary; cervix; prostate; testis, genitourinary
tract; esophagus; larynx, glioblastoma; neuroblastoma; stomach;
skin, keratoacanthoma; lung, epidermoid carcinoma, large cell
carcinoma, small cell carcinoma, lung adenocarcinoma; bone; colon;
colorectal; adenoma; pancreas, adenocarcinoma; thyroid, follicular
carcinoma, undifferentiated carcinoma, papillary carcinoma;
seminoma; melanoma; sarcoma; bladder carcinoma; liver carcinoma and
biliary passages; kidney carcinoma; myeloid disorders; lymphoid
disorders, Hodgkin's, hairy cells; buccal cavity and pharynx
(oral), lip, tongue, mouth, pharynx; small intestine; colon-rectum,
large intestine, rectum, brain and central nervous system; chronic
myeloid leukemia (CML), and leukemia.
In some embodiments, the compounds of this invention are useful for
treating cancer, such as colorectal, thyroid, breast, and lung
cancer; and myeloproliferative disorders, such as polycythemia
vera, thrombocythemia, myeloid metaplasia with myelofibrosis,
chronic myelogenous leukemia, chronic myelomonocytic leukemia,
hypereosinophilic syndrome, juvenile myelomonocytic leukemia, and
systemic mast cell disease.
In some embodiments, the compounds of this invention are useful for
treating hematopoietic disorders, in particular, acute-myelogenous
leukemia (AMLi), chronic-myelogenous leukemia (CML),
acute-promyelocytic leukemia, and acute lymphocytic leukemia
(ALL).
In some embodiments, the compounds of this invention are useful for
treating a cancer or proliferative disease pertinent to gastic
organs, gastrointestinal tract, and digestive organs including the
stomach, small intestine, large intestine, tongue, salivary glands,
pancreas, liver, and gallbladder.
In some embodiments, the compounds of this invention are useful for
treating Barretts' esophagus, esophageal cancer, salivary gland
malignancies, colon and colorectal cancer, intestinal cancer,
gastric cancer, pancreatic cancer, skin cancer and
neuroblastoma.
One aspect of this invention provides compounds that are useful for
the treatment of diseases, disorders, and damages in liver.
In some embodiments, the compounds of this invention are useful for
treating a fatty liver disease, non-alcoholic fatty acid liver
disease (NAFLD), non-alcoholic steatohepatitis (NASH), fatty liver
disease resulting from hepatitis, fatty liver disease resulting
from obesity, fatty liver disease resulting from diabetes, fatty
liver disease resulting from insulin resistance, fatty liver
disease resulting from hypertriglyceridemia, Abetalipoproteinemia,
glycogen storage diseases, Weber-Christian disease, Wolmans
disease, acute fatty liver of pregnancy, or lipodystrophy.
One aspect of this invention provides compounds that are useful for
the treatment of a neurodegenerative disease.
In some embodiments, the disease is Alzheimer's disease (AD),
Parkinson's disease (PD), Huntington's (HD) diseases, amyotrophic
lateral sclerosis (ALS), spinal muscular atrophy (SMA),
schizophrenia, attention-deficit/hyperactivity disorder (ADHD),
fetal alcohol syndrome and diabetic encephalopathy.
Another aspect of this invention provides a method for the
treatment or lessening the severity of a disease selected from a
cancer, a proliferative or hyperproliterative disease, a liver
disease, or a neurodegenerative disease, comprising administering
an effective amount of a compound, or a pharmaceutically acceptable
composition comprising a compound, to a subject in need
thereof.
As inhibitors of protein kinases, the compounds and compositions of
this invention are also useful in biological samples. One aspect of
the invention relates to inhibiting protein kinase activity in a
biological sample, which method comprises contacting said
biological sample with a compound of the invention or a composition
comprising said compound. The term "biological sample", as used
herein, means an in vitro or an ex vivo sample, including, without
limitation, cell cultures or extracts thereof, biopsied material
obtained from a mammal or extracts thereof; and blood, saliva,
urine, feces, semen, tears, or other body fluids or extracts
thereof. Inhibition of protein kinase activity in a biological
sample is useful for a variety of purposes that are known to one of
skill in the art. Examples of such purposes include, but are not
limited to, blood transfusion, organ-transplantation, and
biological specimen storage.
Another aspect of this invention relates to the study of protein
kinases in biological and pathological phenomena; the study of
intracellular signal transduction pathways mediated by such protein
kinases; and the comparative evaluation of new protein kinase
inhibitors. Examples of such uses include, but are not limited to,
biological assays such as enzyme (e.g. kinetics, binding and
inhibition) assays, gel shift assays (electrophoretic mobility
shift assay) and cell-based assays.
The activity of the compounds as protein kinase inhibitors may be
assayed in vitro, in vivo or in a cell line. In vitro assays
include assays that determine inhibition of either the kinase
activity or ATPase activity of the activated kinase. Alternate in
vitro assays quantitate the ability of the inhibitor to bind to the
protein kinase and may be measured either by radiolabelling the
inhibitor prior to binding, isolating the inhibitor/kinase complex
and determining the amount of radiolabel bound, or by running a
competition experiment where new inhibitors are incubated with the
kinase bound to known radioligands. Preferably, in vitro assays
quantitate the ability of the inhibitor to bind to the protein
kinase and may be measured either by probing the inhibitor with
fluorescent molecules prior to binding, isolating the
inhibitor/kinase complex and determining the amount of the bound
probes, utilization of a mobility shift assay with substrates
treated or non-treated with protein kinase, for example, in the
presence of ATP plus inhibitor or DMSO control, or by running a
competition experiment where new inhibitors are incubated with the
kinase bound to known fluorescent probes.
Detailed conditions for assaying a compound utilized in this
invention as an inhibitor of various kinases are set forth in the
Examples below.
In accordance with the foregoing, the present invention further
provides a method for preventing or treating any of the diseases or
disorders described above in a subject in need of such treatment,
which method comprises administering to said subject a
therapeutically effective amount of a compound of the invention or
a pharmaceutically acceptable salt thereof. For any of the above
uses, the required dosage will vary depending on the mode of
administration, the particular condition to be treated and the
effect desired.
Pharmaceutical Compositions
In another aspect, the invention provides a pharmaceutical
composition comprising a kinase inhibitor compound (e.g. DCLK1/2)
as described herein, or a pharmaceutically acceptable ester, salt,
or prodrug thereof, together with a pharmaceutically acceptable
carrier.
Compounds of the invention can be administered as pharmaceutical
compositions by any conventional route, in particular enterally,
e.g., orally, e.g., in the form of tablets or capsules, or
parenterally, e.g., in the form of injectable solutions or
suspensions, topically, e.g., in the form of lotions, gels,
ointments or creams, or in a nasal or suppository form.
Pharmaceutical compositions comprising a compound of the present
invention in free form or in a pharmaceutically acceptable salt
form in association with at least one pharmaceutically acceptable
carrier or diluent can be manufactured in a conventional manner by
mixing, granulating or coating methods. For example, oral
compositions can be tablets or gelatin capsules comprising the
active ingredient together with a) diluents, e.g., lactose,
dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; b)
lubricants, e.g., silica, talcum, stearic acid, its magnesium or
calcium salt and/or polyethyleneglycol; for tablets also c)
binders, e.g., magnesium aluminum silicate, starch paste, gelatin,
tragacanth, methylcellulose, sodium carboxymethylcellulose and or
polyvinylpyrrolidone; if desired d) disintegrants, e.g., starches,
agar, alginic acid or its sodium salt, or effervescent mixtures;
and/or e) absorbents, colorants, flavors and sweeteners. Injectable
compositions can be aqueous isotonic solutions or suspensions, and
suppositories can be prepared from fatty emulsions or suspensions.
The compositions may be sterilized and/or contain adjuvants, such
as preserving, stabilizing, wetting or emulsifying agents, solution
promoters, salts for regulating the osmotic pressure and/or
buffers. In addition, they may also contain other therapeutically
valuable substances. Suitable formulations for transdermal
applications include an effective amount of a compound of the
present invention with a carrier. A carrier can include absorbable
pharmacologically acceptable solvents to assist passage through the
skin of the host. For example, transdermal devices are in the form
of a bandage comprising a backing member, a reservoir containing
the compound optionally with carriers, optionally a rate
controlling barrier to deliver the compound to the skin of the host
at a controlled and predetermined rate over a prolonged period of
time, and means to secure the device to the skin. Matrix
transdermal formulations may also be used. Suitable formulations
for topical application, e.g., to the skin and eyes, are preferably
aqueous solutions, ointments, creams or gels well-known in the art.
Such may contain solubilizers, stabilizers, tonicity enhancing
agents, buffers and preservatives.
Compounds of the invention can be administered in therapeutically
effective amounts in combination with one or more therapeutic
agents (pharmaceutical combinations). For example, synergistic
effects can occur with other immunomodulatory or anti-inflammatory
substances, for example when used in combination with cyclosporin,
rapamycin, or ascomycin, or immunosuppressant analogues thereof,
for example cyclosporin A (CsA), cyclosporin G, FK-506, rapamycin,
or comparable compounds, corticosteroids, cyclophosphamide,
azathioprine, methotrexate, brequinar, leflunomide, mizoribine,
mycophenolic acid, mycophenolate mofetil, 15-deoxyspergualin,
immunosuppressant antibodies, especially monoclonal antibodies for
leukocyte receptors, for example MHC, CD2, CD3, CD4, CD7, CD25,
CD28, B7, CD45, CD58 or their ligands, or other immunomodulatory
compounds, such as CTLA41g. Where the compounds of the invention
are administered in conjunction with other therapies, dosages of
the co-administered compounds will of course vary depending on the
type of co-drug employed, on the specific drug employed, on the
condition being treated and so forth.
The pharmaceutical compositions of the present invention comprise a
therapeutically effective amount of a compound of the present
invention formulated together with one or more pharmaceutically
acceptable carriers. As used herein, the term "pharmaceutically
acceptable carrier" means a non-toxic, inert solid, semi-solid or
liquid filler, diluent, encapsulating material or formulation
auxiliary of any type. The pharmaceutical compositions of this
invention can be administered to humans and other animals orally,
rectally, parenterally, intracisternally, intravaginally,
intraperitoneally, topically (as by powders, ointments, or drops),
buccally, or as an oral or nasal spray.
Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, microemulsions, solutions,
suspensions, syrups and elixirs. In addition to the active
compounds, the liquid dosage forms may contain inert diluents
commonly used in the art such as, for example, water or other
solvents, solubilizing agents and emulsifiers such as ethyl
alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl
alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,
dimethylformamide, oils (in particular, cottonseed, groundnut,
corn, germ, olive, castor, and sesame oils), glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid
esters of sorbitan, and mixtures thereof. Besides inert diluents,
the oral compositions can also include adjuvants such as wetting
agents, emulsifying and suspending agents, sweetening, flavoring,
and perfuming agents.
Injectable preparations, for example, sterile injectable aqueous or
oleaginous suspensions may be formulated according to the known art
using suitable dispersing or wetting agents and suspending agents.
The sterile injectable preparation may also be a sterile injectable
solution, suspension or emulsion in a nontoxic parenterally
acceptable diluent or solvent, for example, as a solution in
1,3-butanediol. Among the acceptable vehicles and solvents that may
be employed are water, Ringer's solution, U.S.P. and isotonic
sodium chloride solution. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium. For this
purpose any bland fixed oil can be employed including synthetic
mono- or diglycerides. In addition, fatty acids such as oleic acid
are used in the preparation of injectables.
In order to prolong the effect of a drug, it is often desirable to
slow the absorption of the drug from subcutaneous or intramuscular
injection. This may be accomplished by the use of a liquid
suspension of crystalline or amorphous material with poor water
solubility. The rate of absorption of the drug then depends upon
its rate of dissolution which, in turn, may depend upon crystal
size and crystalline form. Alternatively, delayed absorption of a
parenterally administered drug form is accomplished by dissolving
or suspending the drug in an oil vehicle.
Compositions for rectal or vaginal administration are preferably
suppositories which can be prepared by mixing the compounds of this
invention with suitable non-irritating excipients or carriers such
as cocoa butter, polyethylene glycol or a suppository wax which are
solid at ambient temperature but liquid at body temperature and
therefore melt in the rectum or vaginal cavity and release the
active compound.
Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like.
The active compounds can also be in micro-encapsulated form with
one or more excipients as noted above. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a magnesium stearate and microcrystalline cellulose. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents.
Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, ear drops, eye
ointments, powders and solutions are also contemplated as being
within the scope of this invention.
The ointments, pastes, creams and gels may contain, in addition to
an active compound of this invention, excipients such as animal and
vegetable fats, oils, waxes, paraffins, starch, tragacanth,
cellulose derivatives, polyethylene glycols, silicones, bentonites,
silicic acid, talc and zinc oxide, or mixtures thereof.
Powders and sprays can contain, in addition to the compounds of
this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or
mixtures of these substances. Sprays can additionally contain
customary propellants such as chlorofluorohydrocarbons.
Transdermal patches have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
can be made by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
According to the methods of treatment of the present invention,
disorders are treated or prevented in a subject, such as a human or
other animal, by administering to the subject a therapeutically
effective amount of a compound of the invention, in such amounts
and for such time as is necessary to achieve the desired result.
The term "therapeutically effective amount" of a compound of the
invention, as used herein, means a sufficient amount of the
compound so as to decrease the symptoms of a disorder in a subject.
As is well understood in the medical arts a therapeutically
effective amount of a compound of this invention will be at a
reasonable benefit/risk ratio applicable to any medical
treatment.
In general, compounds of the invention will be administered in
therapeutically effective amounts via any of the usual and
acceptable modes known in the art, either singly or in combination
with one or more therapeutic agents. A therapeutically effective
amount may vary widely depending on the severity of the disease,
the age and relative health of the subject, the potency of the
compound used and other factors. In general, satisfactory results
are indicated to be obtained systemically at daily dosages of from
about 0.03 to 2.5 mg/kg per body weight. An indicated daily dosage
in the larger mammal, e.g. humans, is in the range from about 0.5
mg to about 100 mg, conveniently administered, e.g. in divided
doses up to four times a day or in retard form. Suitable unit
dosage forms for oral administration comprise from ca. 1 to 50 mg
active ingredient.
In certain embodiments, a therapeutic amount or dose of the
compounds of the present invention may range from about 0.1 mg/Kg
to about 500 mg/Kg, alternatively from about 1 to about 50 mg/Kg.
In general, treatment regimens according to the present invention
comprise administration to a patient in need of such treatment from
about 10 mg to about 1000 mg of the compound(s) of this invention
per day in single or multiple doses. Therapeutic amounts or doses
will also vary depending on route of administration, as well as the
possibility of co-usage with other agents.
Upon improvement of a subject's condition, a maintenance dose of a
compound, composition or combination of this invention may be
administered, if necessary. Subsequently, the dosage or frequency
of administration, or both, may be reduced, as a function of the
symptoms, to a level at which the improved condition is retained
when the symptoms have been alleviated to the desired level,
treatment should cease. The subject may, however, require
intermittent treatment on a long-term basis upon any recurrence of
disease symptoms.
It will be understood, however, that the total daily usage of the
compounds and compositions of the present invention will be decided
by the attending physician within the scope of sound medical
judgment. The specific inhibitory dose for any particular patient
will depend upon a variety of factors including the disorder being
treated and the severity of the disorder; the activity of the
specific compound employed; the specific composition employed; the
age, body weight, general health, sex and diet of the patient; the
time of administration, route of administration, and rate of
excretion of the specific compound employed; the duration of the
treatment; drugs used in combination or coincidental with the
specific compound employed; and like factors well known in the
medical arts.
The invention also provides for a pharmaceutical combinations, e.g.
a kit, comprising a) a first agent which is a kinase inhibitor
compound as disclosed herein, in free form or in pharmaceutically
acceptable salt form, and b) at least one co-agent. The kit can
comprise instructions for its administration.
The terms "co-administration" or "combined administration" or the
like as utilized herein are meant to encompass administration of
the selected therapeutic agents to a single patient, and are
intended to include treatment regimens in which the agents are not
necessarily administered by the same route of administration or at
the same time.
The term "pharmaceutical combination" as used herein means a
product that results from the mixing or combining of more than one
active ingredient and includes both fixed and non-fixed
combinations of the active ingredients. The term "fixed
combination" means that the active ingredients, e.g. a compound of
the invention and a co-agent, are both administered to a patient
simultaneously in the form of a single entity or dosage. The term
"non-fixed combination" means that the active ingredients, e.g. a
compound of the invention and a co-agent, are both administered to
a patient as separate entities either simultaneously, concurrently
or sequentially with no specific time limits, wherein such
administration provides therapeutically effective levels of the two
compounds in the body of the patient. The latter also applies to
cocktail therapy, e.g. the administration of three or more active
ingredients.
In certain embodiments, these compositions optionally further
comprise one or more additional therapeutic agents. For example,
chemotherapeutic agents or other antiproliferative agents may be
combined with the compounds of this invention to treat
proliferative diseases and cancer. Examples of known
chemotherapeutic agents include, but are not limited to,
Gleevec.TM., adriamycin, dexamethasone, vincristine,
cyclophosphamide, fluorouracil, topotecan, taxol, interferons, and
platinum derivatives.
Other examples of agents the compounds of this invention may also
be combined with include, without limitation: treatments for
Alzheimer's Disease such as Aricept18 and Excelon.RTM.; treatments
for Parkinson's Disease such as L-DOPA/carbidopa, entacapone,
ropinrole, pramipexole, bromocriptine, pergolide, trihexephendyl,
and amantadine; agents for treating Multiple Sclerosis (MS) such as
beta interferon (e.g., Avonex.RTM. and Rebif.RTM.), Copaxone.RTM.,
and mitoxantrone; treatments for asthma such as albuterol and
Singulair.RTM.; agents for treating schizophrenia such as zyprexa,
risperdal, seroquel, and haloperidol; anti-inflammatory agents such
as corticosteroids, TNF blockers, IL-I RA, azathioprine,
cyclophosphamide, and sulfasalazine; immunomodulatory and
immunosuppressive agents such as cyclosporin, tacrolimus,
rapamycin, mycophenolate mofetil, interferons, corticosteroids,
cyclophophamide, azathioprine, and sulfasalazine; neurotrophic
factors such as acetylcholinesterase inhibitors, MAO inhibitors,
interferons, anti-convulsants, ion channel blockers, riluzole, and
antiparkinsonian agents; agents for treating cardiovascular disease
such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium
channel blockers, and statins; agents for treating liver disease
such as corticosteroids, cholestyramine, interferons, and
anti-viral agents; agents for treating blood disorders such as
corticosteroids, antileukemic agents, and growth factors; and
agents for treating immunodeficiency disorders such as gamma
globulin. Some examples of materials which can serve as
pharmaceutically acceptable carriers include, but are not limited
to, ion exchangers, alumina, aluminum stearate, lecithin, serum
proteins, such as human serum albumin, buffer substances such as
phosphates, glycine, sorbic acid, or potassium sorbate, partial
glyceride mixtures of saturated vegetable fatty acids, water, salts
or electrolytes, such as protamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, wool fat, sugars such
as lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa
butter and suppository waxes, oils such as peanut oil, cottonseed
oil; safflower oil; sesame oil; olive oil; corn oil and soybean
oil; glycols; such a propylene glycol or polyethylene glycol;
esters such as ethyl oleate and ethyl laurate, agar; buffering
agents such as magnesium hydroxide and aluminum hydroxide; alginic
acid; pyrogen-free water, isotonic saline; Ringer's solution; ethyl
alcohol, and phosphate buffer solutions, as well as other non-toxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition, according
to the judgment of the formulator. The protein kinase inhibitors or
pharmaceutical salts thereof may be formulated into pharmaceutical
compositions for administration to animals or humans. These
pharmaceutical compositions, which comprise an amount of the
protein inhibitor effective to treat or prevent a protein
kinase-mediated condition and a pharmaceutically acceptable
carrier, are another embodiment of the present invention.
In another aspect, the invention provides a kit comprising a
compound capable of inhibiting kinase activity selected from one or
more of the kinase inhibitor compounds described herein, and
instructions for use in treating cancer.
EXAMPLES
The compounds and processes of the present invention will be better
understood in connection with the following examples, which are
intended as an illustration only and not to limit the scope of the
invention. Various changes and modifications to the disclosed
embodiments will be apparent to those skilled in the art and such
changes and modifications including, without limitation, those
relating to the chemical structures, substituents, derivatives,
formulations and/or methods of the invention may be made without
departing from the spirit of the invention and the scope of the
appended claims.
Abbreviations
DCLK1, doublecortin like kinase 1.
DCLK2, doublecortin like kinase 2.
Ni-NTA, nickel-nitrilotriacetic acid
FAM, fluorescein amidite
HTS, high-throughput screening
HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
DTT, dithiothreitol
EDTA, ethylenediaminetetraacetic acid
Chemistry
General Methods
Unless otherwise noted, reagents and solvents were obtained from
commercial suppliers and were used without further purification.
.sup.1H NMR spectra were recorded on a 500 MHz Bruker Avance III
spectrometer and chemical shifts are reported in parts per million
(ppm, .delta.) downfield from tetramethylsilane (TMS). Coupling
constants (J) are reported in Hz. Spin multiplicities are described
as s (singlet), br (broad singlet), d (doublet), t (triplet), q
(quartet), and m (multiplet). Mass spectra were obtained on a
Waters Acquity I UPLC. Preparative HPLC was performed on a Waters
Sunfire C18 column (19 mm.times.50 mm, 5 .mu.M) using a gradient of
15-95% methanol in water containing 0.05% trifluoroacetic acid
(TFA) over 22 min (28 min run time) at a flow rate of 20 mL/min.
Assayed compounds were isolated and tested as TFA salts. Purities
of assayed compounds were in all cases greater than 95%, as
determined by reverse-phase HPLC analysis.
Synthesis
i. Scheme 1: Synthesis of Compound 1
##STR00067##
Compound 1 can be synthesized using the following scheme 1.
##STR00068##
##STR00069##
In Scheme 1, ethyl
2-((2-chloro-5-nitropyrimidin-4-yl)(methyl)amino)benzoate (2) can
be synthesized as follows: A mixture of ethyl
2-(methylamino)benzoate (1.44 g, 8.0 mmol), diisopropylethylamine
(DIEA) (2.8 mL, 16.0 mmol) and 2,4-dichloro-5-nitropyrimidine (2.30
g, 12.0 mmol) in dioxane (40 mL) was heated at 50.degree. C. for 6
hours. After the reaction was complete as monitored by thin layer
chromatography (TLC), the reaction solution was concentrated and
the residue was purified by silica-gel column chromatography with
ethyl acetate and hexane (1/20, v/v) to give the title compound
(2.51 g, 93%). .sup.1H NMR (600 MHz, CDCl3) .delta. 8.44 (s, 1H),
8.02 (d, J=7.2 Hz, 1H), 7.59 (t, J=7.2 Hz, 1H), 7.44 (t, J=7.2 Hz,
1H), 7.22 (d, J=7.8 Hz, 1H), 4.28-4.18 (m, 2H), 3.58 (s, 3H), 1.29
(t, J=7.2 Hz, 3H). .sup.13C NMR (150 MHz, CDCl3) .delta. 164.4,
160.8, 157.0, 155.2, 142.8, 134.1, 132.5, 128.9, 127.7, 61.6, 42.0,
14.0. MS (ESI) m/z 337 (M+H).sup.+
##STR00070##
In Scheme 1,
2-chloro-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-
-one (3) can be synthesized as follows:
A mixture of (2) (2.35 g, 6.98 mmol) and iron power (3.9 g, 69.8
mmol) in acetic acid (100 mL) was heated at 50.degree. C. for 9
hours. After the reaction was complete as monitored by reverse
phase analytical liquid-chromatography electrospray mass
spectrometry (LC-MS), the excess of iron was removed and the
mixture was concentrated in vacuo. The resulting residue was poured
into ice-water which resulted in a solid precipitate that was
collected by filtration, washed with water and air dried to give
the title compound (1.55 g, 85%). .sup.1H NMR (600 MHz, DMSO-d6)
.delta. 10.44 (s, 1H), 8.14 (s, 1H), 7.72 (d, J=4.8 Hz, 1H), 7.58
(s, 1H), 7.27 (d, J=6.0 Hz, 1H), 7.21 (s, 1H), 3.33 (s, 3H).
.sup.13C NMR (150 MHz, DMSO-d6) .delta. 167.6, 161.4, 153.4, 149.7,
147.9, 134.2, 132.0, 125.9, 124.6, 124.3, 120.1, 37.2. MS (ESI) m/z
261 (M+H).sup.+
##STR00071##
In Scheme 1,
2-chloro-5,11-dimethyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazep-
in-6-one (4) can be synthesized as follows:
To a stirred suspension of (3) 688 mg, 2.64 mmol) and Met (0.25 mL,
4.0 mmol) in dimethyl acetamide (DMA, 40.0 mL) was added NaH (360
mg, 60% suspension in mineral oil) at -10.degree. C. and the
reaction was gradually warmed to 0.degree. C. After the reaction
was complete as monitored by LC-MS, the solution was poured into
ice-water which resulted in a solid precipitate. The precipitate
was collected by filtration, washed with water and air dried to
give the title compound (563 mg, 77%). .sup.1H NMR (600 MHz,
DMSO-d6) .delta. 8.57 (s, 1H), 7.68 (dd, J=1.2, 7.2 Hz, 1H),
7.54-7.51 (m, 1H), 7.25 (d, J=7.8 Hz, 1H), 7.20-7.18 (m, 1H), 3.41
(s, 3H), 3.32 (s, 3H). .sup.13C NMR (150 MHz, DMSO-d6) .delta.
167.1, 163.8, 153.7, 153.4, 148.6, 133.5, 132.4, 128.7, 126.0,
124.6, 118.9, 38.1, 36.4. MS (ESI) m/z 275 (M+H).sup.+.
##STR00072##
In Scheme 1,
2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-5,11-dimethyl-5,11--
dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one (XMD8-85-1,
or Compound 1) can be synthesized as follows:
A mixture of (4) (26 mg, 0.1 mmol),
2-methoxy-4-(4-methylpiperizin-1-yl)benzamine (22 mg, 0.1 mmol),
X-Phos (4.3 mg), Pd.sub.2(dba).sub.3 (5.5 mg) and K.sub.2CO.sub.3
(41.5 mg, 0.3 mmol) in 1.2 mL of t-BuOH was heated at 100.degree.
C. in a seal tube for 4 h. Then the reaction was filtered through
celite and eluted with dichloromethane. The dichloromethane was
removed in vacuo and the resulting crude product was purified by
reverse-phase prep-HPLC using a water (0.05% TFA)/acetonitrile
(0.05% TFA) gradient to afford the compounds as TFA salt (20 mg,
yield: 36%). .sup.1H NMR (600 MHz, CD3OD) .delta. 7.83 (dd, J=1.2,
7.8 Hz, 1H), 7.71 (s, 1H), 7.62-7.59 (m, 2H), 7.30 (d, J=8.4 Hz,
1H), 7.26 (t, J=7.2 Hz, 1H), 6.75 (d, J=1.8 Hz, 1H), 6.66 (d, J=7.2
Hz, 1H), 3.92-3.87 (m, 5H), 3.66-3.60 (m, 2H), 3.49 (s, 3H),
3.30-3.24 (m, 2H), 3.14-3.08 (m, 2H), 2.97 (s, 3H). MS (ESI) m/z
446 (M+H).sup.+.
ii. Scheme 2: Synthesis of Compound 2
##STR00073##
Compound 2:
2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-11-methyl-5-(2,2,2--
trifluoroethyl)-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one
(FMF-03-146-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.85 (s, 1H), 8.48 (s,
1H), 8.18 (s, 1H), 7.78-7.67 (m, 2H), 7.57-7.50 (m, 1H), 7.30-7.25
(m, 1H), 7.20 (td, J=7.6, 1.0 Hz, 1H), 6.71 (d, J=2.6 Hz, 1H),
6.61-6.53 (m, 1H), 3.86 (d, J=4.4 Hz, 1H), 3.83 (s, 2H), 3.54 (d,
J=12.0 Hz, 2H), 3.29 (s, 2H), 3.23-3.12 (m, 2H), 2.95 (t, J=12.5
Hz, 2H), 2.88 (s, 3H). MS (ESI) m/z 528 (M+H).sup.+
Compound 2 can be synthesized using the following Scheme 2.
##STR00074##
##STR00075##
In Scheme 2,
2-chloro-11-methyl-5-(2,2,2-trifluoroethyl)-5,11-dihydro-6H-benzo[e]pyrim-
ido[5,4-b][1,4]diazepin-6-one (5) can be synthesized as follows: 1,
2-chloro-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-
-one (3) (500 mg, 1.9 mmol), Cs.sub.2CO.sub.3 (1.3 g, 4.0 mmol) and
trifluoroethyl iodide (250 .mu.L, 2.6 mmol) were dissolved in DMA
(5 mL) and heated to 70.degree. C. for 2 h. The solvent was
evaporated and the residue purified by silica column chromatography
(0-5% MeOH in DCM) to give the title compound (116 mg, 18%).
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 8.84 (s, 1H), 7.73 (dd,
J=7.8, 1.7 Hz, 1H), 7.59 (ddd, J=8.4, 7.3, 1.7 Hz, 1H), 7.34 (dd,
J=8.4, 0.9 Hz, 1H), 7.26 (ddd, J=8.0, 7.3, 1.0 Hz, 1H), 5.04 (br s,
2H), 3.38 (s, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) .delta.
167.79, 165.40, 155.37, 149.08, 134.04, 132.42, 126.15, 125.90,
125.17, 124.92, 123.66, 119.15, 48.61 (q, J=32.5 Hz), 36.30. MS
(ESI) m/z 343 (M+H).sup.+
Other steps in Scheme 2 can be performed as described in Scheme
1.
iii. Scheme 3: Synthesis of Compound 3
##STR00076##
Compound 3:
5-ethyl-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-11-methyl-5-
,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one
(FMF-03-055-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.75 (s, 1H), 8.35 (s,
1H), 7.81 (d, J=8.7 Hz, 1H), 7.64 (dd, J=7.7, 1.7 Hz, 1H), 7.49
(ddd, J=8.7, 7.2, 1.7 Hz, 1H), 7.24-7.13 (m, 2H), 6.72 (d, J=2.6
Hz, 1H), 6.57 (dd, J=8.8, 2.6 Hz, 1H), 4.20-3.92 (m, 1H), 3.89 (s,
2H), 3.83 (s, 3H), 3.54 (d, J=12.1 Hz, 2H), 3.28 (s, 3H), 3.18 (q,
J=10.9 Hz, 2H), 2.94 (t, J=12.6 Hz, 2H), 2.89 (d, J=3.4 Hz, 3H),
1.16 (t, J=7.1 Hz, 3H). MS (ESI) m/z 474 (M+H).sup.+
Compound 3 can be synthesized using the following Scheme 3.
##STR00077##
##STR00078##
In Scheme 3,
2-chloro-5-ethyl-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]di-
azepin-6-one (6) can be synthesized as follows: 1,
2-chloro-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-
-one (3) (500 mg, 1.9 mmol), Cs.sub.2CO.sub.3 (1.3 g, 4.0 mmol) and
ethyl iodide (210 .mu.L, 2.6 mmol) were dissolved in DMA (5 mL) and
heated to 70.degree. C. for 2 h. The solvent was evaporated and the
residue purified by silica column chromatography (0-5% MeOH in DCM)
to give the title compound (280 mg, 51%). .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 8.65 (s, 1H), 7.66 (dd, J=7.8, 1.7 Hz, 1H),
7.53 (ddd, J=8.4, 7.3, 1.7 Hz, 1H), 7.27 (dd, J=8.4, 1.0 Hz, 1H),
7.22 (td, J=7.5, 1.0 Hz, 1H), 4.10 (q, J=5.2 Hz, 2H), 3.18 (s, 3H),
1.20 (t, J=7.1 Hz, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6)
.delta. 167.06, 164.74, 153.97, 153.61, 148.71, 133.30, 132.08,
127.49, 126.71, 124.70, 118.82, 46.09, 36.31, 13.82. MS (ESI) m/z
290 (M+H).sup.+
Other steps in Scheme 3 can be performed as described in Scheme
1.
iv. Scheme 4: Synthesis of Compound 4
##STR00079##
Compound 4 can be synthesized using the following Scheme 4.
##STR00080##
##STR00081##
In Scheme 4,
2-chloro-5-isopropyl-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,-
4]diazepin-6-one (7) can be synthesized as follows: 1,
2-chloro-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-
-one (3) (500 mg, 1.9 mmol), Cs.sub.2CO.sub.3 (1.3 g, 4.0 mmol) and
isopropyl iodide (250 .mu.L, 2.2 mmol) were dissolved in DMA (5 mL)
and heated to 70.degree. C. for 2 h. The solvent was evaporated and
the residue purified by silica column chromatography (0-5% MeOH in
DCM) to give the title compound (167 mg, 29%). .sup.1H NMR (500
MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H), 7.69 (dd, J=7.8, 1.7 Hz,
1H), 7.52 (ddd, J=8.3, 7.3, 1.7 Hz, 1H), 7.27 (dd, J=8.5, 1.1 Hz,
1H), 7.21 (td, J=7.5, 1.0 Hz, 1H), 4.52 (hept, J=6.8 Hz, 1H), 3.35
(s, 3H), 1.80-0.85 (m, 6H). .sup.13C NMR (126 MHz, DMSO-d.sub.6)
.delta. 167.49, 166.18, 154.39, 148.79, 133.11, 132.29, 126.90,
125.90, 124.56, 118.46, 54.22, 36.01. MS (ESI) m/z 304
(M+H).sup.+
Other steps in Scheme 4 can be performed as described in Scheme
1.
v. Other Compounds
All other compounds were prepared from intermediates (4), (5), (6)
or (7) by analogous methods to XMD8-85 (Compound 1) as described
herein.
##STR00082##
Compound 5:
5-ethyl-2-((4-(4-hydroxypiperidin-1-yl)-2-methoxyphenyl)amino)-11-methyl--
5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one
(XMD9-22)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 8.33 (s, 1H), 7.94 (s,
1H), 7.72 (d, J=8.8 Hz, 1H), 7.63 (dd, J=7.7, 1.8 Hz, 1H),
7.50-7.45 (m, 1H), 7.21 (d, J=8.4 Hz, 1H), 7.16 (t, J=7.4 Hz, 1H),
6.61 (d, J=2.6 Hz, 1H), 6.49 (dd, J=8.8, 2.6 Hz, 1H), 4.15-4.01 (m,
1H), 3.80 (s, 3H), 3.62 (tq, J=8.5, 4.4 Hz, 1H), 3.55-3.46 (m, 2H),
3.35 (s, 2H), 3.27 (s, 3H), 1.83 (dq, J=12.4, 4.0 Hz, 2H), 1.50
(dtd, J=13.0, 9.5, 3.8 Hz, 2H), 1.15 (t, J=7.1 Hz, 3H). MS (ESI)
m/z 475 (M+H).sup.+
##STR00083##
Compound 6:
5-isopropyl-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-11-meth-
yl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one
(FMF-03-149-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.71 (s, 1H), 8.25 (s,
1H), 8.11 (s, 1H), 7.78 (d, J=8.7 Hz, 1H), 7.66 (dd, J=7.7, 1.7 Hz,
1H), 7.46 (ddd, J=8.7, 7.2, 1.7 Hz, 1H), 7.27-7.12 (m, 2H), 6.71
(d, J=2.6 Hz, 1H), 6.57 (dd, J=8.8, 2.6 Hz, 1H), 4.60 (hept, J=6.8
Hz, 1H), 3.82 (s, 6H), 3.54 (d, J=12.2 Hz, 2H), 3.28 (s, 3H), 3.17
(dd, J=12.8, 9.2 Hz, 2H), 2.94 (t, J=12.7 Hz, 2H), 2.88 (d, J=3.3
Hz, 3H), 1.49-1.36 (m, 3H), 1.08 (d, J=6.8 Hz, 3H). MS (ESI) m/z
488 (M+H).sup.+
##STR00084##
Compound 7:
4-((5-ethyl-11-methyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b][1,4]d-
iazepin-2-yl)amino)-3-methoxy-N-(1-methylpiperidin-4-yl)benzamide
(FMF-03-059-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.38 (s, 1H), 8.49 (s,
1H), 8.36 (dd, J=8.0, 5.3 Hz, 2H), 8.23-8.16 (m, 1H), 7.66 (dd,
J=7.8, 1.7 Hz, 1H), 7.60-7.54 (m, 1H), 7.54-7.46 (m, 2H), 7.25 (dt,
J=8.4, 1.9 Hz, 1H), 7.19 (td, J=7.5, 1.1 Hz, 1H), 4.61 (br s, 2H),
4.11-3.98 (m, 1H), 3.95 (d, J=3.8 Hz, 3H), 3.49 (d, J=12.2 Hz, 2H),
3.36 (s, 3H), 3.17-3.05 (m, 2H), 2.79 (d, J=4.6 Hz, 3H), 2.08-1.98
(m, 2H), 1.82-1.69 (m, 2H), 1.18 (t, J=7.0 Hz, 3H). MS (ESI) m/z
516 (M+H).sup.+
##STR00085##
Compound 8:
3-methoxy-4-((11-methyl-6-oxo-5-(2,2,2-trifluoroethyl)-6,11-dihydro-5H-be-
nzo[e]pyrimido[5,4-b][1,4]diazepin-2-yl)amino)-N-(1-methylpiperidin-4-yl)b-
enzamide (FMF-03-148-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.43 (s, 1H), 8.63 (s,
1H), 8.38 (d, J=7.5 Hz, 1H), 8.34 (d, J=10.0 Hz, 1H), 8.29 (d,
J=8.4 Hz, 1H), 7.71 (dd, J=7.7, 1.7 Hz, 1H), 7.56 (ddt, J=10.1,
8.7, 3.3 Hz, 2H), 7.53-7.48 (m, 1H), 7.30 (dd, J=8.2, 3.0 Hz, 1H),
7.25-7.20 (m, 1H), 5.27 (s, 1H), 4.62 (s, 1H), 4.17-3.97 (m, 1H),
3.94 (d, J=3.7 Hz, 3H), 3.48 (s, 2H), 3.37 (d, J=2.8 Hz, 3H),
3.18-3.05 (m, 2H), 2.79 (d, J=4.6 Hz, 3H), 2.08-2.01 (m, 2H), 1.78
(qd, J=13.7, 3.9 Hz, 2H).
MS (ESI) m/z 570 (M+H).sup.+
##STR00086##
Compound 9:
4-((5-isopropyl-11-methyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b][1-
,4]diazepin-2-yl)amino)-3-methoxy-N-(1-methylpiperidin-4-yl)benzamide
(FMF-03-151-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.44 (s, 1H), 8.39 (s,
1H), 8.33 (d, J=8.4 Hz, 1H), 8.26 (s, 1H), 7.68 (dd, J=7.7, 1.8 Hz,
1H), 7.57 (dd, J=8.5, 1.9 Hz, 1H), 7.52-7.46 (m, 2H), 7.24 (dt,
J=8.7, 1.9 Hz, 1H), 7.18 (td, J=7.5, 1.0 Hz, 1H), 4.59 (p, J=6.8
Hz, 1H), 4.20-3.98 (m, 1H), 3.94 (d, J=3.9 Hz, 3H), 3.49 (d, J=12.2
Hz, 2H), 3.36 (s, 3H), 3.17-3.06 (m, 2H), 2.79 (d, J=4.6 Hz, 3H),
2.09-1.99 (m, 2H), 1.78 (qd, J=13.6, 3.9 Hz, 2H), 1.47 (s, 3H),
1.11 (s, 3H). MS (ESI) m/z 530 (M+H).sup.+
##STR00087##
Compound 10:
N-(4-((5,11-dimethyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b][1,4]di-
azepin-2-yl)amino)-3-methoxyphenyl)methanesulfonamide
(FMF-03-047-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.57 (s, 1H), 8.36 (s,
1H), 8.16 (s, 1H), 7.98 (d, J=8.6 Hz, 1H), 7.69 (dd, J=7.8, 1.7 Hz,
1H), 7.51 (ddd, J=8.7, 7.2, 1.8 Hz, 1H), 7.26 (d, J=8.3 Hz, 1H),
7.18 (t, J=7.5 Hz, 1H), 6.91 (d, J=2.3 Hz, 1H), 6.84 (dd, J=8.6,
2.3 Hz, 1H), 3.82 (s, 3H), 3.39 (s, 3H), 3.32 (s, 4H), 2.98 (s,
3H). MS (ESI) m/z 455 (M+H).sup.+
##STR00088##
Compound 11:
N-(4-((5-ethyl-11-methyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b][1,-
4]diazepin-2-yl)amino)-3-methoxyphenyl)methanesulfonamide
(FMF-03-087-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.57 (s, 1H), 8.39 (s,
1H), 8.16 (s, 1H), 7.98 (d, J=8.6 Hz, 1H), 7.64 (dd, J=7.7, 1.7 Hz,
1H), 7.53-7.44 (m, 1H), 7.24 (dd, J=8.4, 0.9 Hz, 1H), 7.17 (td,
J=7.5, 1.0 Hz, 1H), 6.91 (d, J=2.4 Hz, 1H), 6.84 (dd, J=8.6, 2.3
Hz, 1H), 4.02 (s, 2H), 3.82 (s, 3H), 3.31 (s, 3H), 2.98 (s, 3H),
1.16 (t, J=7.1 Hz, 3H). MS (ESI) m/z 469 (M+H).sup.+
##STR00089##
Compound 12:
N-(3-methoxy-4-((11-methyl-6-oxo-5-(2,2,2-trifluoroethyl)-6,11-dihydro-5H-
-benzo[e]pyrimido[5,4-b][1,4]diazepin-2-yl)amino)phenyl)methanesulfonamide
(FMF-03-147-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 8.65 (s, 1H), 7.72 (dd,
J=7.8, 1.7 Hz, 1H), 7.60-7.54 (m, 1H), 7.32 (dd, J=8.5, 1.0 Hz,
1H), 7.23 (td, J=7.5, 0.9 Hz, 1H), 6.80 (s, 1H), 6.70 (d, J=1.3 Hz,
2H), 3.75 (s, 3H), 3.65 (s, 2H), 3.29 (s, 3H), 3.17 (s, 3H). MS
(ESI) m/z 523 (M+H).sup.+
##STR00090##
Compound 13:
N-(4-((5-isopropyl-11-methyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b-
][1,4]diazepin-2-yl)amino)-3-methoxyphenyl)methanesulfonamide
(FMF-03-150-2)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.57 (s, 1H), 8.30 (s,
1H), 8.16 (s, 1H), 7.96 (d, J=8.6 Hz, 1H), 7.66 (dd, J=7.8, 1.7 Hz,
1H), 7.50-7.43 (m, 1H), 7.25-7.21 (m, 2H), 7.16 (td, J=7.5, 1.0 Hz,
1H), 6.91 (d, J=2.3 Hz, 1H), 6.83 (dd, J=8.6, 2.3 Hz, 1H), 4.55
(dq, J=38.7, 6.8 Hz, 1H), 3.81 (s, 3H), 3.30 (s, 3H), 2.98 (s, 3H),
1.44 (d, J=5.6 Hz, 4H), 1.09 (d, J=6.6 Hz, 4H).
MS (ESI) m/z 483 (M+H).sup.+
##STR00091##
Compound 14:
5-ethyl-2-((2-methoxy-4-(4-(pyrrolidin-1-yl)piperidine-1-carbonyl)phenyl)-
amino)-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-on-
e (FMF-03-058-2)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.78 (s, 1H), 8.47 (s,
1H), 8.29 (d, J=8.2 Hz, 1H), 8.20 (s, 1H), 7.66 (dd, J=7.7, 1.7 Hz,
1H), 7.50 (ddd, J=8.7, 7.3, 1.8 Hz, 1H), 7.25 (dd, J=8.4, 1.0 Hz,
1H), 7.19 (td, J=7.5, 1.0 Hz, 1H), 7.09-7.00 (m, 2H), 4.05 (s, 2H),
3.91 (s, 3H), 3.53 (s, 2H), 3.47-3.37 (m, 1H), 3.35 (s, 3H),
3.16-3.05 (m, 2H), 2.96 (s, 1H), 2.55 (s, 2H), 2.09 (s, 2H),
2.06-1.92 (m, 2H), 1.93-1.78 (m, 2H), 1.63-1.49 (m, 1H), 1.17 (t,
J=7.1 Hz, 3H).
MS (ESI) m/z 556 (M+H).sup.+
##STR00092##
Compound 15:
N-(4-((5-ethyl-11-methyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b][1,-
4]diazepin-2-yl)amino)phenyl)methanesulfonamide (FMF-03-083-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.66 (s, 1H), 9.44 (s,
1H), 8.43 (s, 1H), 7.74-7.68 (m, 2H), 7.64 (dd, J=7.7, 1.7 Hz, 1H),
7.48 (ddd, J=8.8, 7.2, 1.7 Hz, 1H), 7.26 (dd, J=8.4, 1.0 Hz, 1H),
7.16 (d, J=8.8 Hz, 3H), 4.15-3.59 (m, 2H), 3.35 (s, 3H), 2.93 (s,
3H), 1.17 (t, J=7.0 Hz, 3H).
MS (ESI) m/z 439 (M+H).sup.+
##STR00093##
Compound 16:
5-ethyl-2-((2-methoxy-4-(2-(4-methylpiperazin-1-yl)ethoxy)phenyl)amino)-1-
1-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one
(FMF-03-086-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 8.35 (s, 1H), 8.14 (s,
1H), 7.79 (d, J=8.7 Hz, 1H), 7.64 (dd, J=7.7, 1.7 Hz, 1H), 7.48
(ddd, J=8.7, 7.2, 1.8 Hz, 1H), 7.25-7.12 (m, 2H), 6.66 (d, J=2.7
Hz, 1H), 6.58 (dd, J=8.8, 2.7 Hz, 1H), 4.21 (t, J=5.2 Hz, 2H), 4.05
(s, 2H), 3.81 (s, 3H), 3.41 (s, 4H), 3.27 (s, 3H), 3.17 (s, 4H),
2.82 (s, 3H), 1.16 (t, J=7.1 Hz, 3H).
MS (ESI) m/z 518 (M+H).sup.+
##STR00094##
Compound 17:
4-((5-ethyl-11-methyl-6-oxo-6,11-dihydro-5H-benzo[e]pyrimido[5,4-b][1,4]d-
iazepin-2-yl)amino)benzenesulfonamide (FMF-03-088-1/-2)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 10.07 (s, 1H), 8.51 (s,
1H), 7.95-7.88 (m, 2H), 7.79-7.72 (m, 2H), 7.66 (dd, J=7.8, 1.7 Hz,
1H), 7.53-7.47 (m, 1H), 7.27 (dd, J=8.4, 1.0 Hz, 1H), 7.21-7.15 (m,
3H), 3.53 (s, 2H), 3.39 (s, 3H), 1.18 (t, J=7.1 Hz, 3H). MS (ESI)
m/z 425 (M+H).sup.+
##STR00095##
Compound 18:
N-(2-(dimethylamino)ethyl)-4-((5-ethyl-11-methyl-6-oxo-6,11-dihydro-5H-be-
nzo[e]pyrimido[5,4-b][1,4]diazepin-2-yl)amino)-3-methoxybenzamide
(FMF-03-061-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.41 (s, 1H), 8.64 (t,
J=5.7 Hz, 1H), 8.50 (s, 1H), 8.38 (d, J=8.5 Hz, 1H), 8.23 (s, 1H),
7.66 (dd, J=7.7, 1.7 Hz, 1H), 7.57 (dd, J=8.5, 1.9 Hz, 1H), 7.53
(d, J=2.0 Hz, 1H), 7.50 (ddd, J=8.8, 7.2, 1.7 Hz, 1H), 7.25 (dd,
J=8.4, 1.0 Hz, 1H), 7.19 (td, J=7.5, 1.0 Hz, 1H), 4.09 (s, 2H),
3.94 (s, 3H), 3.62 (q, J=5.9 Hz, 2H), 3.36 (s, 3H), 3.28 (q, J=5.9
Hz, 2H), 2.87 (d, J=4.7 Hz, 6H), 1.18 (t, J=7.0 Hz, 3H). MS (ESI)
m/z 490 (M+H).sup.+
##STR00096##
Compound 19:
2-((4-(3-(dimethylamino)pyrrolidine-1-carbonyl)-2-methoxyphenyl)amino)-5--
ethyl-11-methyl-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one
(FMF-03-067-1)
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 10.17 (s, 1H), 8.48 (s,
1H), 8.32 (d, J=8.2 Hz, 1H), 8.22 (s, 1H), 7.66 (dd, J=7.7, 1.7 Hz,
1H), 7.50 (ddd, J=8.7, 7.3, 1.7 Hz, 1H), 7.27-7.15 (m, 4H), 3.95
(s, 2H), 3.92 (s, 3H), 3.75-3.66 (m, 2H), 3.61 (s, 2H), 3.35 (s,
3H), 2.85 (s, 6H), 2.40-2.26 (m, 1H), 2.12 (t, J=10.6 Hz, 1H), 1.17
(t, J=7.0 Hz, 3H).
MS (ESI) m/z 516 (M+H).sup.+
Selectivity Data
FIG. 1 illustrates the selectivity of Compound 2 (FMF-03-146-1) at
a concentration of 1 .mu.M against a panel of 468 human kinases and
human mutant kinases. These selectivity data were generated using
KINOMEscan.RTM. platform and these images were generated using
TREEspot.TM. Software Tool and reprinted with permission from
KINOMEscan.RTM., a division of DiscoveRx Corporation, .COPYRGT.
DISCOVERX CORPORATION 2010.
Biochemical Assay
General Material
Lysis Buffer
50 mM HEPES pH 7.8
350 mM NaCl
20 mM imidazole
5% Glycerol
Wash Buffer 1--Same as Lysis
Wash Buffer 2 and 3--Same as lysis but 25 mM Imidazole
Elution Buffer--Same as lysis but 300 mM Imidazole
S200 Gel filtration Buffer
10 mM HEPES pH 7.8
700 mM NaCl
1 mM MgCl2
5% Glycerol
Substrate for Gel Shift Assay
5-FAM-KKLRRTLSVA-COOH
DCLK1 Plasmid Construct
The DNA construct consisting of N-terminally 6-His tagged human
DCLK1 residues G351-H689 was prepared. Thus prepared plasmid was
co-transformed with lambda phosphatase under chloramphenicol
selection into BL21(DE3) E. coli cells.
DCLK1 Protein Purification
Protein expression was induced from the DCLK1 plasmid with 0.6 mM
IPTG and expression was allowed to continue for about 10 hours at a
temperature of 18.degree. C. Cells was harvested by centrifugation
and resuspended in Lysis buffer with protease inhibitors (1 mM
Benzamidine and 1 mM PMSF). Lysis was performed by passing 3 times
through a homogenizer. Lysate was centrifuged at 20K for 1 hour at
a temperature of 4.degree. C. and the supernatant was filtered
through a 0.2 micron membrane. Protein was loaded on nickel
(Ni)-NTA resin, washed with Wash buffers, and eluted with 300 mM
imidazole buffer. Eluate was concentrated to 2 mL and passed over a
Superdex 5200 column.
DCLK1 Mobility Shift Assay (Gel Shift Assay)
DCLK1 kinase activity was measured in vitro using an
electrophoretic mobility shift assay. The reaction was assembled in
a 384-well plate in a total volume of 20 .mu.l. The reaction
contained 30 nM recombinant DCLK1, one DCLK1 inhibitor or DMSO, 100
.mu.M ATP and 1 .mu.M FAM-labeled peptide substrate in a buffer
(100 mM HEPES pH 7.5, 0.003% Brij-35, 0.004% Tween-20, 10 mM
MgCl.sub.2, and 2 mM DTT). DCLK1 inhibitors were dispensed using a
Labcyte Echo liquid handler. The reaction was incubated at room
temperature for two hours and quenched by addition of 40 .mu.L of
termination buffer (100 mM HEPES pH 7.3, 0.015% Brij-35, 0.1% CR-3,
1.times.CR-8, and 40 mM EDTA). Substrate and product peptides
present in each sample were electrophoretically separated and
detected using 12-channel LabChip3000 microfluidic capillary
electrophoresis instrument (Caliper Life Sciences, Waltham Mass.,
USA). The change in the relative fluorescence intensities of
substrate and product peaks (reflecting enzyme activity) was
measured. Capillary electrophoregrams were analyzed using HTS Well
Analyzer software (Caliper Life Sciences, Waltham Mass., USA).
The kinase activity in each sample was determined as the
product-to-sum ratio (PSR):P/(S+P), where P is the peak height of
the product peptide and S is the peak height of the substrate
peptide. Negative control samples (DMSO in the absence of
inhibitor) and positive control samples (100% inhibition, a tested
DCLK1 inhibitor) were assembled in replicates and were used to
calculate percent inhibition values for each compound at each
concentration.
Percent inhibition (% Inhibition) was determined using the
following equation:
.times..times..times..times..times..times. ##EQU00001## where
PSR.sub.inh is the product-sum ratio in the presence of inhibitor,
PSR.sub.0% is the average product-sum ration in the absence of
inhibitor and PSR.sub.100% is the average product-sum ratio in
100%-inhibition control samples.
The DCLK1 candidate inhibitors were tested in 8-point dose-response
format on each assay plate. The IC.sub.50 values were determined by
fitting the inhibition curves by an eight dose-response model using
GraphPad Prism 7 software.
FIG. 2 shows the thus obtained IC.sub.50 values of Compound 2,
Compound 12, Compound 8, Compound 6, Compound 13 and Compound 9. In
particular, the IC.sub.50 values of Compound 2 and Compound 8 were
substantially decreased from the IC.sub.50 value of the positive
control.
INCORPORATION BY REFERENCE
The contents of all references (including literature references,
issued patents, published patent applications, and co-pending
patent applications) cited throughout this application are hereby
expressly incorporated herein in their entireties by reference.
Unless otherwise defined, all technical and scientific terms used
herein are accorded the meaning commonly known to one with ordinary
skill in the art.
EQUIVALENTS
Those skilled in the art will recognize, or be able to ascertain
using no more than routine experimentation, many equivalents of the
specific embodiments of the invention described herein. Such
equivalents are intended with be encompassed by the following
claims.
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